Biomarkers for lupus

ABSTRACT

The present invention provides biomarkers valuable in diagnosing or detecting a susceptibility to lupus. The present invention also provides panels of biomarkers valuable in diagnosing or detecting a susceptibility to lupus with good specificity and selectivity. Said biomarkers and panels of biomarkers are also useful in monitoring the progression or regression of lupus, or monitoring the progression to a flare of the disease or transition from a flare into remission or monitoring the efficacy of a therapeutic agent to lupus.

FIELD OF THE INVENTION

The present invention relates to novel biomarkers for lupus and methods and kits for their use. Also provided are vaccines for the treatment or prevention of lupus.

BACKGROUND

Systemic lupus erythematosus (SLE) or lupus is a chronic autoimmune disease that can affect the joints and almost every major organ in the body, including heart, kidneys, skin, lungs, blood vessels, liver, and the nervous system. As in other autoimmune diseases, the body's immune system attacks the body's own tissues and organs, leading to inflammation. A person's risk to develop lupus appears to be determined mainly by genetic factors, but environmental factors, such as infection or stress may trigger the onset of the disease. The course of lupus varies, and is often characterised by alternating periods of flares, i.e. increased disease activity, and periods of remission.

It has been estimated that in the US, about 1 million people suffer from lupus, world-wide, conservative estimates indicate about 4 million lupus patients. Lupus occurs about 9 times more frequently in women than in men. Although lupus can occur at any age, it is most common in women of childbearing age.

There is presently no cure for lupus. However, there are means of reducing and dealing with flares, including drugs such as anti-inflammatory drugs, alternative medicines or life-style changes. The treatment regimes will depend on the severity of the disease, and the responsiveness of the patient. Disease-modifying antirheumatic drugs can be used preventively to reduce the incidence of flares. When flares occur, they are often treated with corticosteroids.

As mentioned above, lupus patients can present with a variety of diverse symptoms, and is therefore difficult to diagnose. A number of tests are used in order to make a diagnosis, including antinuclear antibody test (ANA), tests for other auto-antibodies such as anti-DNA antibodies, tests for serum complement levels, urine analysis, looking for elevated protein levels in urine, and sometimes biopsies of an affected organ are used. However, these tests do not necessarily give a definitive diagnosis; for example, a positive ANA test can occur due to infections or rheumatic diseases, and even healthy people without lupus can test positive. The sensitivity and specificity for the ANA test are 93% and 57% (Habash-Bseiso,D Clin Med Res. 2005 August; 3(3): 190-193). Lupus diagnosis often takes 7 to 10 years using current methods.

There is therefore a need for new or improved biomarkers for lupus that can be used in methods of diagnosing lupus, for the early detection of lupus, subclinical or presymptomatic lupus or a predisposition to lupus, or for monitoring the progression of lupus or the likelihood to transition from remission to flare or vice versa, or the efficacy of a therapeutic treatment thereof.

SUMMARY OF THE INVENTION

It has now been found that the presence of certain auto-antibodies or combinations of auto-antibodies in a patient, or a significant increase in the number or concentration of such auto-antibodies or combinations of auto-antibodies provide useful biomarkers for lupus. Early results indicate that the disclosed invention will also distinguish lupus from other diseases.

Therefore the present invention provides biomarkers valuable in diagnosing or detecting a susceptibility to lupus. The present invention also provides panels of biomarkers valuable in diagnosing lupus or detecting a susceptibility to lupus. The present disclosure includes panels with good specificity and/or selectivity as compared to prior art methods. Said biomarkers and panels of biomarkers are also useful in monitoring the progression or regression of lupus, or monitoring the progression to a flare of the disease or transition from a flare into remission or monitoring the efficacy of a therapeutic agent to lupus.

In one embodiment, the present invention provides methods/uses and products (e.g. biomarkers, kits, vaccines, panels and methods of using the same) which employ ZMAT2 (including variants thereof (see discussion below)) as an auto-antigen, optionally with at least 1, 2, 3, 4, 5, 7, 9, 10, 15 or 20 other auto-antigens (e.g. as recited in Tables 1, 2, 3, 4 or 5) also being used.

Suitably the at least 1, 2, 3, 4, 5, 7, 9, 10, 15, 20 or 25 other auto-antigens that are used are selected from the group consisting of:

-   -   (a) BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding         protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein         5); FUS (Fusion (involved in t(12;16) in malignant         liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B         (High-mobility group 20B); HOXB6 (Homeo box B6, transcript         variant 2); KRT8 (Keratin 8); LIN28 (Lin-28 homolog (C.         elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein         3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1);         PHLDA1 (Pleckstrin homology-like domain, family A, member 1);         PIAS2 (Msx-interacting-zinc finger, transcript variant alpha);         RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN         (RAN, member RAS oncogene family); RARA (Retinoic acid receptor         alpha); RBMS1 (RNA binding motif, single stranded interacting         protein 1, transcript variant); RDBP (RD RNA binding protein);         RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN         and FYVE domain containing 1); SMN1 (Survival of motor neuron 1,         telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's         syndrome nuclear autoantigen 1); STAU. (Staufen, RNA binding         protein (Drosophila), transcript variant T3); STK11         (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1         (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone         MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant         5); and ZNF38 (Zinc finger protein 38); or     -   (b) ASPSCR1 (alveolar soft part sarcoma chromosome region,         candidate 1);

CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); KRT8 (Keratin 8); L1N28 Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 inducible kinase); SRPK1 (SFRS protein kinase 1); STAU1 (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); ZMAT2 (Zinc finger, matrin type 2); and ZNF38. (Zinc finger protein 38).

In addition to ZMAT2 and the at least 1, 2, 3, 4, 5, 7, 9, 10, 15, 20 or 25 other auto-antigens (e.g. as selected from group (a) or (b) above), one or more further auto-antigens may be employed wherein said one or more further auto-antigens are selected from the group consisting of: IFI16 (Interferon, gamma-inducible protein 16); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki), transc); RALBP1 (RalA-binding protein 1); SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)); and BANK1 (B-cell scaffold protein with ankyrin repeats 1).

Thus, it will be appreciated that the methods of the invention may comprise testing a patient sample against ZMAT2/a combination of auto-antigens which combination may be as outlined above (including variants thereof). In this way, the presence, absence or a modulated level of auto-antibodies in the sample may be detected. In some embodiments the auto-antigens are provided/used in the form of a panel and methods of using these panels as well as the panels themselves form part of the present invention.

Examples of particular methods/uses and products employing ZMAT2/a combination of auto-antigens as outlined above will be apparent from the description and the appended claim set of this application. Also, specific examples of methods/uses and products as described herein may be modified so as to employ any particular combination of auto-antigens that can be derived from the above discussion of the methods/uses and products of the present invention.

The methods, biomarkers, kits and panels of the present invention may be used alone or in combination with (e.g. before, after or concurrently with) one or more other diagnostic/prognostic indicators or techniques. Such a combination may for instance be used to to: (i) diagnose lupus; (ii) detect a susceptibility to lupus; (iii) monitor the progression or regression of lupus; (iv) monitor the progression to a flare of the disease or transition from a flare into remission; or (v) monitor the efficacy of a therapeutic agent to lupus.

Examples of diagnostic/prognostic indicators or techniques which may be employed in combination with the disclosure of the present invention can include those which have utility in the diagnosis/prognosis of lupus. Examples may include: the presence of symptoms characteristic or indicative of lupus; a complete or incomplete SLEDAI (Systemic Lupus Erythematosus Disease Activity Index) score; anti-nuclear antibody (ANA) blood test; tests for other auto-antibodies such as anti-DNA antibodies; tests for serum complement levels; urine analysis (e.g. looking for elevated protein levels in urine); the use of one or more other biomarkers; anti-ds DNA test; blood counts; and biopsy analysis of an affected organ.

The present invention provides a method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises:

(a) the auto-antigens listed in Table 2,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 6, 7, 8, 9 or 10 ore more auto-antigens are deleted or replaced.

As will be appreciated from the definition of “diagnosis” (see below), the above method may be useful in providing a positive or negative diagnosis of lupus (or a predisposition thereto).

The auto-antigens of Table 2 are ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); BANK1 B-cell (scaffold protein with ankyrin repeats 1); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); IFI16 (Interferon, gamma-inducible protein 16); KRT8 (Keratin 8); L1N28 Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki)); RALBP1 (RalA binding protein 1); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1); SSA2 (Sjogren syndrome antigen A2 (6010a, ribonucleoprotein autoantigen SS-A/Ro)); STAU1 (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); ZNF38 (Zinc finger protein 38).

The present invention provides a method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises:

(a) the auto-antigens listed in Table 1,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced.

The auto-antigens of Table 1 are ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); IFI16 (Interferon, gamma-inducible protein 16); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 KDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RALBP1 (RalA-binding protein 1); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); ZNF38 (Zinc finger protein 38), PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki), transc), and SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)).

The present invention provides a method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises:

(a) the auto-antigens listed in Table 4,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced.

The auto-antigens of Table 4 are ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 or MAP1S (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); IFI16 (Interferon, gamma-inducible protein 16); KRT8 (Keratin 8); LIN28 (Lin-28 homolog (C. elegans)); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki)); RALBP1 (RalA binding protein 1); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); SRPK1 (SFRS protein kinase 1); SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); ZNF38 (Zinc finger protein 38).

The present invention provides a method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises:

(a) the auto-antigens listed in Table 3,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 10, 15, 20 or 25 or more auto-antigens are deleted or replaced.

The auto-antigens of Table 3 are ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 or MAP1S (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); IFI16 (Interferon, gamma-inducible protein 16); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki)); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RALBP1 (RalA binding protein 1); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor, alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1) ;SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)); SSNA1 Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); VCL (Vinculin); ZNF38 (Zinc finger protein 38).

The present invention also provides a method of diagnosing a patient as having or being predisposed to developing lupus, comprising detecting in a sample taken from said patient one or more auto-antibodies that bind an auto-antigen selected from the group consisting of the auto-antigens listed in Table 3 (e.g. ZMAT2), wherein the presence of said one or more auto-antibodies, or an increase in the concentration of said one or more auto-antibodies, indicates that the patient suffers from or is predisposed to develop lupus.

The present invention further provides a method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises at least 10 auto-antigens selected from Table 3 or from Table 1 or from Table 2.

Preferably the panel comprises at least 15 or 20 auto-antigens and/or preferably the panel comprises ZMAT2.

In one embodiment the at least 10, 15 or 20 auto-antigens are selected from the group consisting of:

-   -   (i) ZMAT2 (Zinc finger, matrin type 2); BPY2IP1 (MAP1S protein);         CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5         (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in         t(12;16) in malignant liposarcoma)); HAGH         (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group         20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin         8); LIN28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH         dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1         (Poly(A) binding protein, cytoplasmic 1); PHLDA 1 (Pleckstrin         homology-like domain, family A, member 1); PIAS2         (Msx-interacting-zinc finger, transcript variant alpha);         RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN         (RAN, member RAS oncogene family); RARA (Retinoic acid receptor         alpha); RBMS1 (RNA binding motif, single stranded interacting         protein 1, transcript variant); RDBP (RD RNA binding protein);         RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN         and FYVE domain containing 1); SMN1 (Survival of motor neuron 1,         telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's         syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding         protein (Drosophila), transcript variant T3); STK11         (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1         (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone         MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant         5); and ZNF38 (Zinc finger protein 38); or     -   (ii) ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft         part sarcoma chromosome region, candidate 1); CEBPG         (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD         (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z         (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS         (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B         (High-mobility group 20B); KRT8 (Keratin 8); LIN28 Lin-28         homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S         (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic         1); PHLDA1 (Pleckstrin homology-like domain, family A, member         1); PIAS2 (Msx-interacting-zinc finger, transcript variant         alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA         (protein kinase, interferon-inducible double stranded RNA         dependent activator); RARA (Retinoic acid receptor, alpha); RDBP         (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31         (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1         (SFRS protein kinase 1); STAU1 (Staufen, RNA binding protein         (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like,         clone MGC:12349); VCL (vinculin); and ZNF38 (Zinc finger protein         38).

Conveniently in the methods of the present invention detecting the presence or an increased level of at least two auto-antibodies provides information useful for diagnosis, preferably detecting the presence of an increased level of at least three, preferably at least four and more preferably at least five auto-antibodies provides information useful for diagnosis.

Advantageously the sample from the patient used in the methods of the present invention is a body fluid, optionally selected from blood, serum, plasma, saliva, lymphatic fluid, wound secretion, urine, faeces, mucus or cerebrospinal fluid (CSF). Typically, the sample may be serum or plasma.

A further aspect of the present invention provides a method of monitoring the progression or regression of lupus, or monitoring the progression to a flare of the disease or transition from a flare into remission, comprising (a) performing methods described above, repeating step (a) using a second sample obtained from the patient at a time later than the first sample; and (c) comparing the auto-antibodies detected in said first and second samples; wherein a change in the level or presence or absence of the auto-antibodies detected between the first and second samples indicates the progression or regression of lupus, or the progression to a flare, or transition from a flare into remission.

Another aspect of the present invention provides a method of monitoring the efficacy in a patient of a therapeutic agent to lupus, comprising (a) performing methods described above (b) administering the therapeutic agent to the patient; (c) performing again method described above using a second sample from the patient taken after the administration of the therapeutic agent; and (d) comparing the auto-antibodies detected in said first and second samples; wherein a change in the level or presence or absence of the auto-antibodies detected in the first and second sample is indicative of the efficacy of the therapeutic agent to lupus.

The present invention also provides a panel of auto-antigens as used in any of the methods described herein. In one embodiment, the present invention provides a panel of auto-antigens, wherein said panel comprises:

(a) the auto-antigens listed in Table 2,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 6, 7, 8, 9 or 10 ore more auto-antigens are deleted or replaced;

(ii) (a) the auto-antigens listed in Table 1,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced;

(iii) (a) the auto-antigens listed in Table 4,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced; or

(iv) (a) the auto-antigens listed in Table 3,

(b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or

(c) the panel of (a) in which up to 10, 15, 20 or 25 or more auto-antigens are deleted or replaced.

As will be appreciated from the description of the invention, other panels of auto-antigens have also been described for use in the methods of the present invention and these panels may also form part of the present invention. In a preferred embodiment, a panel of the invention comprises ZMAT2.

In one embodiment a panel of the invention is for use in: (i) diagnosing lupus or determining the severity thereof; (ii) detecting a susceptibility to lupus; (iii) monitoring the progression or regression of lupus; (iv) monitoring the progression to a flare of the disease or transition from a flare into remission; or (v) monitoring the efficacy of a therapeutic agent to lupus.

FIGURES

FIG. 1 is a cartoon illustrating a method for the detection of auto-antibodies in serum.

DESCRIPTION

As explained further in the Examples below, in order to elucidate biomarkers valuable in diagnosing or detecting susceptibility to lupus, the inventors developed microarrays of antigens suitable for detecting antibodies in samples. Subsequently, a large number of body fluid samples obtained from individuals positively diagnosed as having lupus with existing methods were analysed for the presence of auto-antibodies binding to auto-antigens immobilized on microarrays. Data analysis of the results obtained allowed detection of useful biomarkers for lupus.

A first data analysis methodology allowed detection of a set of 35 biomarkers, auto-antigens to which the patient serum samples contained auto-antibodies, and which are set out in Table 1 below.

A second data analysis methodology allowed detection of a set of 32 biomarkers which are set out in Table 2 below.

Between the two analysis methods a total of 45 useful biomarkers were identified which are set out in Table 3 below. The first and second analysis methods identified 22 biomarkers in common These shared biomarkers are set out in Table 4 below. The biomarkers which were unique to each set are set out in Table 5 below.

In one embodiment of the invention, modified versions of Tables 1, 2, 3, 4 or 5 are envisaged in which 1, 2, 3, 4 or all 5 of the following markers are removed from the Table: IFI16 (Interferon, gamma-inducible protein 16); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki), transc); RALBP1 (RalA-binding protein 1); SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)); and BANK1 (B-cell scaffold protein with ankyrin repeats 1).

In such a scenario a reference to a panel comprising, say, 5, auto-antigens of Table 1 would be a reference to a panel comprising 5 auto-antigens selected from the group consisting of: ZMAT2 (Zinc finger, matrin type 2); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11 FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); ZNF38 (Zinc finger protein 38).

Likewise, in such a scenario, a reference to a panel comprising, say, 5, auto-antigens of Table 2 would be a reference to a panel comprising 5 auto-antigens selected from the group consisting of: ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); KRT8 (Keratin 8); L1N28 Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1); STAU1 (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); ZNF38 (Zinc finger protein 38).

According to one aspect of the present invention therefore there is provided a method of diagnosing a patient as having or being predisposed to developing lupus, said method comprising detecting in a sample taken from said patient one or more auto-antibodies that bind one or more auto-antigens; wherein the presence of said one or more auto-antibodies, or an increase in the concentration of said one or more auto-antibodies, indicates that the patient suffers or is predisposed to develop lupus, wherein said one or more auto-antigens are selected from ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11 FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); or ZNF38 (Zinc finger protein 38).

Preferably, the method comprises detecting auto-antibodies to one or more of the auto-antigens listed above, in combination with one or more of the auto-antigens IFI16 (Interferon, gamma-inducible protein 16); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki), transc), SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)), and RALBP1 (RalA-binding protein 1).

The sample may be a body fluid, and is preferably selected from blood, serum, plasma saliva, lymphatic fluid, wound secretion, urine, faeces, mucus and cerebrospinal fluid (CSF). Typically, the sample may be serum or plasma.

The term “diagnosis” (and for the avoidance of doubt, grammatical variants thereof such as “diagnosing”, “diagnostics” etc.) as used herein is intended to be construed broadly. Thus, for example, the methods and products of the present invention are envisaged as being useful in the following: making a positive or negative diagnosis of lupus in a patient (including subclinical or presymptomatic lupus); assessing the severity of lupus in a patient; assessing an individual's susceptibility to lupus; providing a prognosis of an individual suffering from lupus; monitoring the progression or regression of lupus; monitoring the progression to a flare of the disease or transition from a flare into remission; or monitoring the efficacy of a therapeutic agent to lupus. As indicated above, the methods and products (kits, biomarkers, panels etc.) of the present invention may be used alone to achieve any of the foregoing or may alternatively be combined with one or more other diagnostic/prognostic indicators (e.g. the identification of one or more symptoms characteristic of lupus, the results of one or more biochemical tests etc.). Also, given the difficulties encountered in diagnosing lupus, it should be appreciated that the term “diagnosis” does not necessarily entail an unequivocal diagnosis of lupus in a patient but may rather, for example, provide an indication of the likelihood of lupus being present or absent in an individual.

By a “patient” (the term is used interchangeably herein with the term “individual”) is meant an individual from whom a sample has been obtained e.g. for testing in accordance with a method of the present invention. The term “patient” is not intended to imply that the individual is necessarily undergoing any treatment and thus the term may refer to any individual who is being tested in accordance with a protocol of the present invention.

By “bind” or “bound” used herein is meant a specific interaction strong enough to withstand standard washing procedures in typical protein binding assays.

Accordingly, the present invention comprehends the use of one or more auto-antibodies that bind to said one or more auto-antigens as biomarkers for lupus.

By an increase in the concentration of said one or more auto-antibodies is meant a statistically significant increase in the concentration of the one or more auto-antibodies in said sample taken from the patient as compared with a predetermined mean concentration found in non-diseased, normal patients. A surrogate normal sample can also be used, for example a pooled normal control measured in relation to normal and diseased samples. Typically, the mean concentration of auto-antibodies found in normal patients may be calculated by reference to samples taken from a population of non-diseased individuals or from pooled samples from such individuals. In some embodiments, an increase of more than 5%, more suitably 10%, and suitably more than 15% or 20%, as compared with such mean may be taken as statistically significant. It will be appreciated that the units are arbitrary. In the case of clinical drug evaluation, there will be a change in auto-antibody concentration upon drug treatment; it is anticipated that the levels of most of these auto-antibodies may decrease post drug treatment.

Suitably, said method may comprise the detection of auto-antibodies that bind two or more of said auto-antigens, preferably 3 or more of said auto-antigens, even more preferably 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or 31 of these auto-antigens. In some embodiments, the patient may be pre-symptomatic.

In another aspect of the present invention there is provided a method of diagnosing a patient as having or being predisposed to developing lupus, said method comprising detecting in a sample from said patient auto-antibodies to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more, even more preferably to a panel comprising all 35, auto-antigens selected from the group consisting of those auto-antigens listed in Table 1 below; wherein the presence or separately a significant increase in the concentration of said auto-antibodies in the sample to substantially all members of the panel indicates that the patient suffers or is predisposed to develop lupus. In some embodiments, said panel may comprise additional auto-antigens (or other antigens) to those specified.

A significant increase in the concentration of the auto-antibodies means a statistically significant increase as determined by methods for statistically analysing experimental results of the kind commonly used in the art. By “substantially all members of the panel” is meant that the sample contains auto-antibodies, or an elevated concentration of auto-antibodies, to at least 50%, preferably at least 75% or 80% and typically at least 90% or 95%, of the auto-antigens included in the panel. It is envisaged that in some embodiments, the presence of auto-antibodies to one or more members of the panel and an increase in the respective concentrations of auto-antibodies to one or more other members of the panel will be indicative of lupus, it being unnecessary in most cases for there to be an increased concentration for all auto-antibodies detected. However, in some cases, the mere presence of auto-antibodies to substantially all members of the panel will be significant.

A significant decrease in the concentration of the auto-antibodies means a statistically significant decrease as determined by methods for statistically analysing experimental results of the kind commonly used in the art. By “substantially all members of the panel” is meant that the sample contains auto-antibodies, or a reduced concentration of auto-antibodies, to at least 50%, preferably at least 75% or 80% and typically at least 90% or 95%, of the auto-antigens included in the panel. It is envisaged that in some embodiments, the presence of auto-antibodies to one or more members of the panel and a decrease in the respective concentrations of auto-antibodies to one or more other members of the panel will be indicative of lupus, it being unnecessary in most cases for there to be a decreased concentration for all auto-antibodies detected.

It is expected that such a panel of biomarkers according to the present invention will provide greatly improved sensitivity or improved specificity in such methods of diagnosis as compared with known biomarkers for lupus.

A panel is defined as a collection of two or more individual markers where the information provided by the panel produces a greater sensitivity and/or specificity than that provided by the markers individually.

As mentioned above, the auto-antigens for lupus are listed in the Tables below (e.g. Table 1). References to gene sequences and uniprot accession numbers for such auto-antigens are also given. However, it will be appreciated that variants of the sequences given, for example allelic variants, orthologues from other mammalian species or other variants (e.g. fragments or antigens with post-translational modifications) that would be bound by the respective auto-antibodies may also be used. Accordingly, it will be understood that the use of such variants are contemplated as being within the scope of the present invention and that references to particular auto-antigens or biomarkers should be construed accordingly. In particular, the present invention comprehends the use of auto-antigens being encoded from respective gene sequences that individually have at least an 80% identity to the corresponding sequences listed in the Tables below (e.g. Table 1). Suitably, such sequences have at least an 85% or 90% identity to the corresponding sequences in the Tables, and preferably they have at least a 95% identity to such sequences, e.g., more than a 96%, 97%, 98% or 99% identity. The skilled person will appreciate that such variants should retain the ability to be bound by the auto-antibodies in a positive patient sample which auto-antibodies recognize the wild-type antigen.

TABLE 1 Gene symbol Name Synonyms Entrez Gene ID and protein sequence link ZMAT2 Zinc finger, FLJ31121 153527; http://ca.expasy.org/uniprot/Q96NC0 matrin type 2 BANK1 B-cell scaffold BANK; 55024; http://ca.expasy.org/uniprot/Q8NDB2 protein with FLJ20706; ankyrin repeats 1 FLJ34204 BPY2IP1 MAP1S protein MAP8; C19orf5; 55201; http://www.ncbi.nlm.nih.gov/entrez/ VCY2IP1; viewer.fcgi?db=protein&val=AAH06358.2 FLJ10669; VCY2IP-1; MGC133087 CEBPG CCAAT/enhancer 1054; http://ca.expasy.org/uniprot/P53567 binding protein (C/EBP), gamma E1B-AP5 E1B-55 kDa- E1BAP5; E1B- 11100; http://ca.expasy.org/uniprot/O76022 associated AP5; HNRPUL1; protein 5 FLJ12944 FUS Fusion (involved TLS; CHOP; 2521; http://ca.expasy.org/uniprot/P35637 in t(12;16) in FUS1; FUS- malignant CHOP; liposarcoma) TLS/CHOP; hnRNP-P2 HAGH Hydroxyacylglutathione GLO2; GLX2; 3029; http://ca.expasy.org/uniprot/Q16775 hydrolase GLXII; HAGH1 HMG20B High-mobility SOXL; HMGX2; 10362; http://ca.expasy.org/uniprot/Q9P0W2 group 20B BRAF25; BRAF35; PP7706; pp8857; FLJ26127; SMARCE1r HOXB6 Homeo box B6, HOX2; HU-2; 3216; http://ca.expasy.org/uniprot/P17509 transcript variant 2 HOX2B; Hox-2.2 IFI16 Interferon, PYHIN2; 3428; http://ca.expasy.org/uniprot/Q16666 gamma-inducible IFNGIP1; protein 16 MGC9466 KRT8 Keratin 8 K8; KO; CK8; 3856; http://ca.expasy.org/uniprot/P05787 CYK8; K2C8; CARD2 LIN28 Lin-28 homolog CSDD1; LIN-28; 79727; http://ca.expasy.org/uniprot/Q9H9Z2 (C. elegans) LIN28A; ZCCHC1; FLJ12457 NDUFV3 NADH CI-9KD 4731; http://ca.expasy.org/uniprot/P56181 dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa PABPC1 Poly(A) binding PAB1; PABP; 26986; http://ca.expasy.org/uniprot/P11940 protein, PABP1; cytoplasmic 1 PABPC2; PABPL1 PHLDA1 Pleckstrin PHRIP; TDAG51; 22822; http://ca.expasy.org/uniprot/Q8WV24 homology-like DT1P1B11; domain, family A, MGC131738 member 1 PIAS2 Msx-interacting- miz; MIZ1; SIZ2; 9063; http://ca.expasy.org/uniprot/Q13105 zinc finger, PIASX; ZMIZ4; transcript variant MGC102682; alpha PIASX-BETA; PIASX-ALPHA PSME3 Proteasome Ki; PA28G; REG- 10197; http://ca.expasy.org/uniprot/Q12920 (prosome, GAMMA; PA28- macropain) gamma activator subunit 3 (PA28 gamma; Ki), transc RAB11FIP3 RAB11 family KIAA0665; 9727; http://ca.expasy.org/uniprot/O75154 interacting Rab11-FIP3 protein 3 (class II) RALBP1 RalA binding RIP; RIP1; 10928; http://ca.expasy.org/uniprot/Q15311 protein 1 RLIP76 RAN RAN, member TC4; Gsp1; 5901; http://ca.expasy.org/uniprot/P17080 RAS oncogene ARA24 family, RARA Retinoic acid RAR; NR1B1 5914; http://ca.expasy.org/uniprot/P10276 receptor, alpha RBMS1 RNA binding YC1; MSSP; 5937; http://ca.expasy.org/uniprot/P29558 motif, single SCR2; MSSP-1; stranded MSSP-2; MSSP- interacting 3; MGC3331; protein 1, MGC15146 transcript variant RDBP RD RNA binding RD; RDP; 7936; http://ca.expasy.org/uniprot/P18615 protein D6S45; NELF-E RNF12 Ring finger RLIM; 51132; http://ca.expasy.org/uniprot/Q9NVW2 protein 12, MGC15161; NY- transcript variant 1 REN-43 RUFY1 RUN and FYVE RABIP4; 80230; http://ca.expasy.org/uniprot/Q96T51 domain ZFYVE12; containing 1 FLJ22251 SMN1 Survival of motor SMA; SMN; 6606; http://ca.expasy.org/uniprot/Q16637 neuron 1, SMA1; SMA2; telomeric SMA3; SMA4; SMA@; SMNT; BCD541; T- BCD541 SRPK1 SFRS protein SFRSK1 6732; http://ca.expasy.org/uniprot/Q96SB4 kinase 1 SSA2 Sjogren RO60; SSA2; 6738; http://ca.expasy.org/uniprot/P10155 syndrome TROVE2 antigen A2 (60 kDa, ribonucleoprotein autoantigen SS- A/Ro) SSNA1 Sjogren's N14; NA14; NA- 8636; http://ca.expasy.org/uniprot/O43805 syndrome 14 nuclear autoantigen 1 STAU Staufen, RNA STAU; FLJ25010 6780; http://ca.expasy.org/uniprot/O95793 binding protein (Drosophila), transcript variant T3 STK11 Serine/threonine PJS; LKB1 6794; http://ca.expasy.org/uniprot/Q15831 kinase 11 (Peutz- Jeghers syndrome) TOM1 Target of myb1 FLJ33404 10043; http://ca.expasy.org/uniprot/O60784 (chicken) TXNL2 Thioredoxin-like, GRX3; GRX4; 10539; http://ca.expasy.org/uniprot/O76003 clone GLRX4; PICOT; MGC: 12349 TXNL2; TXNL3; FLJ11864; bA500G10.4; GLRX3 TXNRD1 Thioredoxin TR; TR1; TXNR; 7296; http://ca.expasy.org/uniprot/Q16881 reductase 1, TRXR1; GRIM- transcript variant 5 12; MGC9145 ZNF38 Zinc finger ZNF38; Zipro1; 7589; http://ca.expasy.org/uniprot/Q9Y5A6 protein 38 NY-REN-21; DKFZp434L134; DKFZp686H10254

The term “sequence identity” refers to the degree to which two polynucleotide or polypeptide sequences are identical on a residue-by-residue basis over a particular region of comparison. The term “percentage of sequence identity” is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleotide base or amino acid occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The identity between two DNA or protein sequences may be determined using computer programs known in the art, such as GAP software provided in the GCG program package with default parameters (Needleman and Wunsch 1970 J Mol Biol 48: 443-453). The identity between two protein sequences is also displayed after searching a protein database with Blast2 (Altschul et al (1997) Nucleic Acids Res. 25 (17) 3389-3402).

It is anticipated that the methods of the present invention may provide greater sensitivity and/or specificity for the diagnosis of lupus than previously available in the art.

By “sensitivity” is meant the proportion of persons with the disease phenotype who test positive.

By “specificity” is meant the proportion of persons without the disease phenotype who test negative.

Advantageously, the methods of the invention may have a sensitivity of >67%, preferably >70%, even more preferably >75%, 80%, 85%, 90%, and most preferably >95%, or a specificity of >85%, preferably >90%, even more preferably >95%, and most preferably >98%.

According to yet another aspect of the invention there is provided a method of monitoring the progression or regression of lupus in a patient, said method comprising detecting in a first sample obtained from a patient one or more auto-antibodies that bind one or more auto-antigens selected from the group consisting of ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); or ZNF38 (Zinc finger protein 38), or auto-antibodies that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more, even more preferably to a panel comprising all 35, auto-antigens selected from the group consisting of those auto-antigens listed in Table 1; detecting said auto-antibodies in a second sample obtained from the patient at a time later than the first sample; wherein a change in the auto-antibodies detected in the first and second samples is indicative of the progression or regression of lupus.

According to yet another aspect of the invention there is provided a method of monitoring the progression towards a flare of the disease, or the transition from a flare to remission, of lupus in a patient, said method comprising detecting in a first sample obtained from a patient one or more auto-antibodies that bind one or more auto-antigens selected from the group consisting of ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK1_(Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); or ZNF38 (Zinc finger protein 38), or auto-antibodies that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more, even more preferably to a panel comprising all 35, auto-antigens selected from the group consisting of those auto-antigens listed in Table 1; detecting said auto-antibodies in a second sample obtained from the patient at a time later than the first sample; wherein a change in the auto-antibodies detected in the first and second samples is indicative of the progression towards a flare, or transition from a flare to a period of remission of the disease.

A change in the auto-antibodies may be a change in the number of different auto-antigens to which auto-antibodies are detected in the first as compared to the second sample, or it may be a change in the concentration of an auto-antibody in the first as compared to the second sample, or it may be a combination of both.

Whilst the biomarkers specifically disclosed herein in relation to the present invention are elevated in lupus patients (and therefore an increased auto-antibody concentration of the biomarkers are indicative of lupus), auto-antibodies for which a decrease in concentration indicates that a patient suffers from or is presymptomatic or predisposed to develop lupus are also envisaged, and may also provide useful biomarkers for lupus. Such markers may be used in combination with the methods or products of the present invention so that an increase in certain biomarkers is indicative of lupus whilst a decrease in certain other biomarkers is also indicative of lupus.

According to yet another aspect of the present invention there is provided a method of monitoring the efficacy of a therapeutic agent to lupus, comprising detecting in a first sample obtained from a patient the presence of one or more auto-antibodies that bind one or more auto-antigens selected from the group consisting of ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11_(Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); or ZNF38 (Zinc finger protein 38), or auto-antibodies that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more, even more preferably to a panel comprising all 35, auto-antigens selected from the group consisting of those auto-antigens listed in Table 1; administering the therapeutic agent to the patient; detecting said one or more auto-antibodies in a second sample taken from the patient taken after the administration of the therapeutic agent; and comparing the one or more auto-antibodies in said first and second samples respectively, wherein a change in the auto-antibodies in the first and second sample is indicative of the efficacy of the therapeutic agent to lupus.

The first sample may be taken prior to the first administration of a therapeutic agent, or after treatment has already commenced. The method can be used to monitor the efficacy of treatment over time, e.g., over days, weeks, months or years.

The therapeutic agent may be any therapeutic agent that can be beneficial for the treatment of lupus, for example anti-rheumatic agents or anti-inflammatory agents.

According to yet another aspect of the present invention, there is provided a method of predicting whether a patient suffering from lupus will respond to treatment with a therapeutic agent, comprising detecting the presence or absence of one or more auto-antibodies that bind to one or more auto-antigens selected from the group consisting of ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11_(Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); or ZNF38 (Zinc finger protein 38), or auto-antibodies that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more, even more preferably to a panel comprising all 35, auto-antigens selected from the group consisting of those auto-antigens listed in Table 1; wherein the presence or absence of said one or more auto-antibodies indicates that the patient will or will not respond to treatment with said therapeutic agent.

The auto-antibodies may be detected by any means known in the art. This includes the use of protein arrays, bead-based assays, Western blots, immunoprecipitation, silver staining, dot blots, etc. The binding of the auto-antibodies can be detected by any means, including enzyme-linked assays such as ELISAs, radioimmunoassays (RIAs), DELFIA assays, fluorescence-based detection methods, bead assays such as Luminex bead assays, etc.

In some embodiments, arrays of said auto-antigens may be employed for detecting said auto-antibodies, for example arrays prepared according to the methods disclosed by WO-A-01/57198 or WO-A-2003/064656; preferred detection methods are fluorescence-based detection methods.

The patient may have or be predisposed to develop the disease naturally. However, a patient may also develop lupus as a result of environmental factors, for example as a side effect of treatment with therapeutic agents or as a result of exposure to toxic materials, or as a result of manipulation, e.g., in animal models used in pharmaceutical research and development. The patient may be a human being, but can also be a non-human animal such, for example, as a mouse, rat, guinea pig, cat, dog, horse, pig, cow, non-human primate (e.g. monkey), or any other animal used for research and drug testing.

Each of the aspects and features of the invention and methods as described above are applicable to the use of auto-antigens from Tables 1, 2, 3, 4 and 5.

Therefore according to the present invention there is provided a method of diagnosing a patient as having or being predisposed to developing lupus, said method comprising detecting in a sample taken from said patient one or more auto-antibodies that bind one or more auto-antigens; wherein the presence of said one or more auto-antibodies, or an increase in the concentration of said one or more auto-antibodies, indicates that the patient suffers from or is predisposed to develop lupus, wherein said one or more antigens are selected from the auto-antigens set out in Table 2.

TABLE 2 Gene Entrez Gene ID and symbol Name Synonyms protein sequence link ZMAT2 Zinc finger, matrin type 2 FLJ31121 153527; http://ca.expasy.org/uniprot/Q96NC0 ASPSCR1 alveolar soft part TUG; ASPL; ASPS; RCC17; 79058; sarcoma chromosome UBXD9; UBXN9; ASPCR1; http://ca.expasy.org/uniprot/Q7Z6N7 region, candidate 1 FLJ45380; ASPSCR1 BANK1 B-cell scaffold protein BANK; FLJ20706; FLJ34204 55024; with ankyrin repeats 1 http://ca.expasy.org/uniprot/Q8NDB2 CEBPG CCAAT/enhancer GPE1BP; IG/EBP-1; CEBPG 1054; binding protein http://ca.expasy.org/uniprot/P53567 (C/EBP), gamma DDX55 DEAD (Asp-Glu-Ala- FLJ16577; KIAA1595; 57696; Asp) box polypeptide MGC33209; DDX55 http://ca.expasy.org/uniprot/Q8NHQ9 55 DOM3Z dom-3 homolog Z (C. elegans) NG6; DOM3L; DOM3Z 1797; http://ca.expasy.org/uniprot/O77932 E1B-AP5 E1B-55 kDa-associated E1BAP5; E1B-AP5; 11100; protein 5 HNRPUL1; FLJ12944 http://ca.expasy.org/uniprot/O76022 FUS Fusion (involved in TLS; CHOP; FUS1; FUS- 2521; t(12;16) in malignant CHOP; TLS/CHOP; hnRNP- http://ca.expasy.org/uniprot/P35637 liposarcoma) P2 HMG20B High-mobility group SOXL; HMGX2; BRAF25; 10362; 20B BRAF35; PP7706; pp8857; http://ca.expasy.org/uniprot/Q9P0W2 FLJ26127; SMARCE1r IFI16 Interferon, gamma- PYHIN2; IFNGIP1; MGC9466 3428; inducible protein 16 http://ca.expasy.org/uniprot/Q16666 KRT8 Keratin 8 K8; KO; CK8; CYK8; K2C8; 3856; CARD2 http://ca.expasy.org/uniprot/P05787 LIN28 Lin-28 homolog (C. elegans) CSDD1; LIN-28; LIN28A; 79727; ZCCHC1; FLJ12457 http://ca.expasy.org/uniprot/Q9H9Z2 LNX ligand of numb-protein X LNX; MPDZ; PDZRN2; LNX1 84708; http://ca.expasy.org/uniprot/Q8TBB1 MAP1S MAP1S protein MAP8; C19orf5; VCY2IP1; 55201; FLJ10669; VCY2IP-1; http://ca.expasy.org/uniprot/Q66K74 MGC133087 PABPC1 Poly(A) binding protein, PAB1; PABP; PABP1; 26986; cytoplasmic 1 PABPC2; PABPL1 http://ca.expasy.org/uniprot/P11940 PHLDA1 Pleckstrin homology- PHRIP; TDAG51; DT1P1B11; 22822; like domain, family A, MGC131738 http://ca.expasy.org/uniprot/Q8WV24 member 1 PIAS2 Msx-interacting-zinc miz; MIZ1; SIZ2; PIASX; 9063; finger, transcript variant ZMIZ4; MGC102682; PIASX- http://ca.expasy.org/uniprot/Q13105 alpha BETA; PIASX-ALPHA PRKCBP1 protein kinase C RACK7; PRKCBP1; 23613; binding protein 1 PRO2893; MGC31836; http://ca.expasy.org/uniprot/Q9H2G5 ZMYND8 PRKRA protein kinase, RAX; PACT; DYT16; HSD14; 8575; interferon-inducible PRKRA http://ca.expasy.org/uniprot/O75569 double stranded RNA dependent activator PSME3 Proteasome (prosome, Ki; PA28G; REG-GAMMA; 10197; macropain) activator PA28-gamma http://ca.expasy.org/uniprot/Q12920 subunit 3 (PA28 gamma; Ki) RALBP1 RalA binding protein 1 RIP; RIP1; RLIP76 10928; http://ca.expasy.org/uniprot/Q15311 RARA Retinoic acid receptor, RAR; NR1B1 5914; alpha http://ca.expasy.org/uniprot/P10276 RDBP RD RNA binding RD; RDP; D6S45; NELF-E 7936; protein http://ca.expasy.org/uniprot/P18615 RPL30 ribosomal protein L30 RPL30 6156; http://ca.expasy.org/uniprot/P04645 RPL31 ribosomal protein L31 MGC88191; RPL31 6160; http://ca.expasy.org/uniprot/P62899 SNK serum-inducible kinase SNK; PLK2 10769; http://ca.expasy.org/uniprot/Q9NYY3 SRPK1 SFRS protein kinase 1 SFRSK1 6732; http://ca.expasy.org/uniprot/Q96SB4 SSA2 Sjogren syndrome RO60; SSA2; TROVE2 6738; antigen A2 (60 kDa, http://ca.expasy.org/uniprot/P10155 ribonucleoprotein autoantigen SS-A/Ro) STAU1 Staufen, RNA binding STAU; FLJ25010 6780; protein (Drosophila), http://ca.expasy.org/uniprot/O95793 transcript variant T3 TXNL2 Thioredoxin-like, clone GRX3; GRX4; GLRX4; 10539; MGC: 12349 PICOT; TXNL2; TXNL3; http://ca.expasy.org/uniprot/O76003 FLJ11864; bA500G10.4; GLRX3 VCL vinculin MVCL; CMD1W; VCL 7414; http://ca.expasy.org/uniprot/P18206 ZNF38 Zinc finger protein 38 ZNF38; Zipro1; NY-REN-21; 7589; DKFZp434L134; http://ca.expasy.org/uniprot/Q9Y5A6 DKFZp686H10254

In another aspect of the present invention provides a method of diagnosing a patient as having or being predisposed to developing lupus, said method comprising detecting in a sample from said patient auto-antibodies to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more, even more preferably to a panel comprising all 32 auto-antigens selected from the group of auto-antigens listed in Table 2.

According to yet another aspect of the invention there is provided a method of monitoring the progression or regression of lupus in a patient, said method comprising detecting in a first sample obtained from a patient one or more auto-antibodies that bind one or more auto-antigens selected from the group of auto-antigens set out in Table 2, or auto-antigens that bind to a panel comprising 5 or more, preferably, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more or even more preferably to a panel comprising all 32, auto-antigens selected from the group of auto-antigens listed in Table 2.

According to yet another aspect of the present invention there is provided a method of monitoring the efficacy of the therapeutic agent lupus, comprising detecting in a first sample obtained from a patient the presence of one or more auto-antibodies that bind to one or more auto-antigens selected from the group consisting of the auto-antigens in Table 2 or to antigens that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 or more or even more preferably to a panel comprising all 32, auto-antigens selected from the group of auto-antigens listed in Table 2.

According to yet another aspect of the present invention there is provided a method of predicting whether a patient suffering from lupus will respond to treatment with a therapeutic agent comprising detecting the presence or absence of one or more auto-antibodies that bind to one or more auto-antigens selected from the group consisting of the auto-antigens set out in Table 2, or auto-antibodies that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more, or preferably to a panel comprising all 32 auto-antigens selected from those auto-antigens listed in Table 2.

As explained above the combined sets of 35 and 32 biomarkers produces 45 biomarkers which have been found to be useful in detecting lupus. These biomarkers are set out in Table 3 below.

TABLE 3 Gene Entrez Gene ID and symbol Name Synonyms protein sequence link ZMAT2 Zinc finger, matrin type 2 FLJ31121 153527; http://ca.expasy.org/uniprot/Q96NC0 ASPSCR1 alveolar soft part sarcoma TUG; ASPL; ASPS; RCC17; 79058; chromosome region, UBXD9; UBXN9; ASPCR1; http://ca.expasy.org/uniprot/Q7Z6N7 candidate 1 FLJ45380; ASPSCR1 BANK1 B-cell scaffold protein with BANK; FLJ20706; FLJ34204 55024; ankyrin repeats 1 http://ca.expasy.org/uniprot/Q8NDB2 BPY2IP1 MAP1S protein MAP8; C19orf5; VCY2IP1; 55201; FLJ10669; VCY2IP-1; http://ca.expasy.org/uniprot/Q66K74 MGC133087 CEBPG CCAAT/enhancer binding GPE1BP; IG/EBP-1; CEBPG 1054; protein (C/EBP), gamma http://ca.expasy.org/uniprot/P53567 DDX55 DEAD (Asp-Glu-Ala-Asp) FLJ16577; KIAA1595; 57696; box polypeptide 55 MGC33209; DDX55 http://ca.expasy.org/uniprot/Q8NHQ9 DOM3Z dom-3 homolog Z (C. elegans) NG6; DOM3L; DOM3Z 1797; http://ca.expasy.org/uniprot/O77932 E1B-AP5 E1B-55 kDa-associated E1BAP5; E1B-AP5; 11100; protein 5 HNRPUL1; FLJ12944 http://ca.expasy.org/uniprot/O76022 FUS Fusion (involved in TLS; CHOP; FUS1; FUS- 2521; t(12;16) in malignant CHOP; TLS/CHOP; hnRNP- http://ca.expasy.org/uniprot/P35637 liposarcoma) P2 HAGH Hydroxyacylglutathione GLO2; GLX2; GLXII; HAGH1 3029; hydrolase http://ca.expasy.org/uniprot/Q16775 HMG20B High-mobility group 20B SOXL; HMGX2; BRAF25; 10362; BRAF35; PP7706; pp8857; http://ca.expasy.org/uniprot/Q9P0W2 FLJ26127; SMARCE1r HOXB6 Homeo box B6, transcript HOX2; HU-2; HOX2B; Hox- 3216; variant 2 2.2 http://ca.expasy.org/uniprot/P17509 IFI16 Interferon, gamma- PYHIN2; IFNGIP1; MGC9466 3428; inducible protein 16 http://ca.expasy.org/uniprot/Q16666 KRT8 Keratin 8 K8; KO; CK8; CYK8; K2C8; 3856; CARD2 http://ca.expasy.org/uniprot/P05787 LIN28 Lin-28 homolog (C. elegans) CSDD1; LIN-28; LIN28A; 79727; ZCCHC1; FLJ12457 http://ca.expasy.org/uniprot/Q9H9Z2 LNX ligand of numb-protein X LNX; MPDZ; PDZRN2; LNX1 84708; http://ca.expasy.org/uniprot/Q8TBB1 NDUFV3 NADH dehydrogenase CI-9KD 4731; (ubiquinone) flavoprotein http://ca.expasy.org/uniprot/P56181 3, 10 kDa PABPC1 Poly(A) binding protein, PAB1; PABP; PABP1; 26986; cytoplasmic 1 PABPC2; PABPL1 http://ca.expasy.org/uniprot/P11940 PHLDA1 Pleckstrin homology-like PHRIP; TDAG51; DT1P1B11; 22822; domain, family A, member 1 MGC131738 http://ca.expasy.org/uniprot/Q8WV24 PIAS2 Msx-interacting-zinc miz; MIZ1; SIZ2; PIASX; 9063; finger, transcript variant ZMIZ4; MGC102682; PIASX- http://ca.expasy.org/uniprot/Q13105 alpha BETA; PIASX-ALPHA PRKCBP1 protein kinase C binding RACK7; PRKCBP1; 23613; protein 1 PRO2893; MGC31836; http://ca.expasy.org/uniprot/Q9H2G5 ZMYND8 PRKRA protein kinase, interferon- RAX; PACT; DYT16; HSD14; 8575; inducible double stranded PRKRA http://ca.expasy.org/uniprot/O75569 RNA dependent activator PSME3 Proteasome (prosome, Ki; PA28G; REG-GAMMA; 10197; macropain) activator PA28-gamma http://ca.expasy.org/uniprot/Q12920 subunit 3 (PA28 gamma; Ki) RAB11FIP3 RAB11 family interacting KIAA0665; Rab11-FIP3 9727; protein 3 (class II) http://ca.expasy.org/uniprot/O75154 RALBP1 RalA binding protein 1 RIP; RIP1; RLIP76 10928; http://ca.expasy.org/uniprot/Q15311 RAN RAN, member RAS TC4; Gsp1; ARA24 5901; oncogene family http://ca.expasy.org/uniprot/P17080 RARA Retinoic acid receptor, RAR; NR1B1 5914; alpha http://ca.expasy.org/uniprot/P10276 RBMS1 RNA binding motif, single YC1; MSSP; SCR2; MSSP-1; 5937; stranded interacting MSSP-2; MSSP-3; http://ca.expasy.org/uniprot/P29558 protein 1, transcript MGC3331; MGC15146 variant RDBP RD RNA binding protein RD; RDP; D6S45; NELF-E 7936; http://ca.expasy.org/uniprot/P18615 RNF12 Ring finger protein 12, RLIM; MGC15161; NY-REN- 51132; transcript variant 1 43 http://ca.expasy.org/uniprot/Q9NVW2 RPL30 ribosomal protein L30 RPL30 6156; http://ca.expasy.org/uniprot/P04645 RPL31 ribosomal protein L31 MGC88191; RPL31 6160; http://ca.expasy.org/uniprot/P62899 RUFY1 RUN and FYVE domain RABIP4; ZFYVE12; 80230; containing 1 FLJ22251 http://ca.expasy.org/uniprot/Q96T51 SMN1 Survival of motor neuron SMA; SMN; SMA1; SMA2; 6606; 1, telomeric SMA3; SMA4; SMA@; http://ca.expasy.org/uniprot/Q16637 SMNT; BCD541; T-BCD541 SNK serum-inducible kinase SNK; PLK2 10769; http://ca.expasy.org/uniprot/Q9NYY3 SRPK1 SFRS protein kinase 1 SFRSK1 6732; http://ca.expasy.org/uniprot/Q96SB4 SSA2 Sjogren syndrome RO60; SSA2; TROVE2 6738; antigen A2 (60 kDa, http://ca.expasy.org/uniprot/P10155 ribonucleoprotein autoantigen SS-A/Ro) SSNA1 Sjogren's syndrome N14; NA14; NA-14 8636; nuclear autoantigen 1 http://ca.expasy.org/uniprot/O43805 STAU Staufen, RNA binding STAU; FLJ25010 6780; protein (Drosophila), http://ca.expasy.org/uniprot/O95793 transcript variant T3 STK11 Serine/threonine kinase PJS; LKB1 6794; 11 (Peutz-Jeghers http://ca.expasy.org/uniprot/Q15831 syndrome) TOM1 Target of myb1 (chicken) FLJ33404 10043; http://ca.expasy.org/uniprot/O60784 TXNL2 Thioredoxin-like, clone GRX3; GRX4; GLRX4; 10539; MGC: 12349 PICOT; TXNL2; TXNL3; http://ca.expasy.org/uniprot/O76003 FLJ11864; bA500G10.4; GLRX3 TXNRD1 Thioredoxin reductase 1, TR; TR1; TXNR; TRXR1; 7296; transcript variant 5 GRIM-12; MGC9145 http://ca.expasy.org/uniprot/Q16881 VCL vinculin MVCL; CMD1W; VCL 7414; http://ca.expasy.org/uniprot/P18206 ZNF38 Zinc finger protein 38 ZNF38; Zipro1; NY-REN-21; 7589; DKFZp434L134; http://ca.expasy.org/uniprot/Q9Y5A6 DKFZp686H10254

Therefore according to the present invention there is provided a method of diagnosing a patient as having or being predisposed to developing lupus, said method comprising detecting in a sample taken from said patient one or more auto-antibodies that bind one or more auto-antigens; wherein the presence of said one or more auto-antibodies, or an increase in the concentration of said one or more auto-antibodies, indicates that the patient suffers or is predisposed to develop lupus, wherein said one or more antigens are selected from the auto-antigens set out in Table 3.

In another aspect of the present invention there is provided a method of diagnosing a patient as having or being predisposed to developing lupus, said method comprising detecting in a sample from said patient auto-antibodies to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or more, even more preferably to a panel comprising all 45 auto-antigens selected from the group consisting of those auto-antigens listed in Table 3.

According to yet another aspect of the invention there is provided a method of monitoring the progression or regression of lupus in a patient, said method comprising detecting in a first sample obtained from a patient one or more auto-antibodies that bind one or more auto-antigens selected from the group of auto-antigens set out in Table 2, or auto-antigens that bind to a panel comprising 5 or more, preferably, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 or more, auto-antigens selected from the auto-antigens listed in Table 3.

According to yet another aspect of the present invention there is provided a method of monitoring the efficacy of the therapeutic agent lupus, comprising detecting in a first sample obtained from a patient the presence of one or more auto-antibodies that bind to one or more auto-antigens selected from the group consisting of the auto-antigens in Table 3 or to antigens that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or more or even more preferably to a panel comprising all 45 auto-antigens selected from the auto-antigens listed in Table 3.

According to yet another aspect of the present invention there is provided a method of predicting whether a patient suffering from lupus will respond to treatment with a therapeutic agent comprising detecting the presence or absence of one or more auto-antibodies that bind to one or more auto-antigens selected from the group consisting of the auto-antigens set out in Table 2, or auto-antibodies that bind to a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or more or preferably to a panel comprising all 45 auto-antigens selected from those auto-antigens listed in Table 3.

Comparison of the set 35 biomarkers in Table 1 with the set of 32 biomarkers in Table 2 reveals a group of shared biomarkers or biomarkers in common.

The shared biomarkers are set out in Table 4, whilst the unshared biomarkers, or those biomarkers unique to each analysis methodology, are set out in Table 5.

TABLE 4 Gene Entrez Gene ID and symbol Name Synonyms protein sequence link ZMAT2 Zinc finger, matrin type 2 FLJ31121 153527; http://ca.expasy.org/uniprot/Q96NC0 BANK1 B-cell scaffold protein with BANK; FLJ20706; FLJ34204 55024; ankyrin repeats 1 http://ca.expasy.org/uniprot/Q8NDB2 BPY2IP1 MAP1S protein MAP8; C19orf5; VCY2IP1; 55201; MAP1S FLJ10669; VCY2IP-1; http://ca.expasy.org/uniprot/Q66K74 MGC133087 CEBPG CCAAT/enhancer binding GPE1BP; IG/EBP-1; CEBPG 1054; protein (C/EBP), gamma http://ca.expasy.org/uniprot/P53567 E1B-AP5 E1B-55 kDa-associated E1BAP5; E1B-AP5; 11100; protein 5 HNRPUL1; FLJ12944 http://ca.expasy.org/uniprot/O76022 FUS Fusion (involved in t(12;16) TLS; CHOP; FUS1; FUS- 2521; in malignant liposarcoma) CHOP; TLS/CHOP; hnRNP- http://ca.expasy.org/uniprot/P35637 P2 HMG20B High-mobility group 20B SOXL; HMGX2; BRAF25; 10362; BRAF35; PP7706; pp8857; http://ca.expasy.org/uniprot/Q9P0W2 FLJ26127; SMARCE1r IFI16 Interferon, gamma-inducible PYHIN2; IFNGIP1; MGC9466 3428; protein 16 http://ca.expasy.org/uniprot/Q16666 KRT8 Keratin 8 K8; KO; CK8; CYK8; K2C8; 3856; CARD2 http://ca.expasy.org/uniprot/P05787 LIN28 Lin-28 homolog (C. elegans) CSDD1; LIN-28; LIN28A; 79727; ZCCHC1; FLJ12457 http://ca.expasy.org/uniprot/Q9H9Z2 PABPC1 Poly(A) binding protein, PAB1; PABP; PABP1; 26986; cytoplasmic 1 PABPC2; PABPL1 http://ca.expasy.org/uniprot/P11940 PHLDA1 Pleckstrin homology-like PHRIP; TDAG51; DT1P1B11; 22822; domain, family A, member 1 MGC131738 http://ca.expasy.org/uniprot/Q8WV24 PIAS2 Msx-interacting-zinc finger, miz; MIZ1; SIZ2; PIASX; 9063; transcript variant alpha ZMIZ4; MGC102682; PIASX- http://ca.expasy.org/uniprot/Q13105 BETA; PIASX-ALPHA PSME3 Proteasome (prosome, Ki; PA28G; REG-GAMMA; 10197; macropain) activator PA28-gamma http://ca.expasy.org/uniprot/Q12920 subunit 3 (PA28 gamma; Ki) RALBP1 RalA binding protein 1 RIP; RIP1; RLIP76 10928; http://ca.expasy.org/uniprot/Q15311 RARA Retinoic acid receptor, RAR; NR1B1 5914; alpha http://ca.expasy.org/uniprot/P10276 RDBP RD RNA binding protein RD; RDP; D6S45; NELF-E 7936; http://ca.expasy.org/uniprot/P18615 SRPK1 SFRS protein kinase 1 SFRSK1 6732; http://ca.expasy.org/uniprot/Q96SB4 SSA2 Sjogren syndrome antigen RO60; SSA2; TROVE2 6738; A2 (60 kDa, http://ca.expasy.org/uniprot/P10155 ribonucleoprotein autoantigen SS-A/Ro) STAU Staufen, RNA binding STAU; FLJ25010 6780; protein (Drosophila), http://ca.expasy.org/uniprot/O95793 transcript variant T3 TXNL2 Thioredoxin-like, clone GRX3; GRX4; GLRX4; 10539; MGC: 12349 PICOT; TXNL2; TXNL3; http://ca.expasy.org/uniprot/O76003 FLJ11864; bA500G10.4; GLRX3 ZNF38 Zinc finger protein 38 ZNF38; Zipro1; NY-REN-21; 7589; DKFZp434L134; http://ca.expasy.org/uniprot/Q9Y5A6 DKFZp686H10254

TABLE 5 Gene Entrez Gene ID and symbol Name Synonyms protein sequence link ASPSCR1 alveolar soft part sarcoma TUG; ASPL; ASPS; RCC17; 79058; chromosome region, UBXD9; UBXN9; ASPCR1; http://ca.expasy.org/uniprot/Q7Z6N7 candidate 1 FLJ45380; ASPSCR1 DDX55 DEAD (Asp-Glu-Ala-Asp) FLJ16577; KIAA1595; 57696; box polypeptide 55 MGC33209; DDX55 http://ca.expasy.org/uniprot/Q8NHQ9 DOM3Z dom-3 homolog Z (C. elegans) NG6; DOM3L; DOM3Z 1797; http://ca.expasy.org/uniprot/O77932 HAGH Hydroxyacylglutathione GLO2; GLX2; GLXII; HAGH1 3029; hydrolase http://ca.expasy.org/uniprot/Q16775 HOXB6 Homeo box B6, transcript HOX2; HU-2; HOX2B; Hox- 3216; variant 2 2.2 http://ca.expasy.org/uniprot/P17509 LNX ligand of numb-protein X LNX; MPDZ; PDZRN2; LNX1 84708; http://ca.expasy.org/uniprot/Q8TBB1 NDUFV3 NADH dehydrogenase CI-9KD 4731; (ubiquinone) flavoprotein http://ca.expasy.org/uniprot/P56181 3, 10 kDa PRKCBP1 protein kinase C binding RACK7; PRKCBP1; 23613; protein 1 PRO2893; MGC31836; http://ca.expasy.org/uniprot/Q9H2G5 ZMYND8 PRKRA protein kinase, interferon- RAX; PACT; DYT16; HSD14; 8575; inducible double stranded PRKRA http://ca.expasy.org/uniprot/O75569 RNA dependent activator RAB11FIP3 RAB11 family interacting KIAA0665; Rab11-FIP3 9727; protein 3 (class II) http://ca.expasy.org/uniprot/O75154 RAN RAN, member RAS TC4; Gsp1; ARA24 5901; oncogene family http://ca.expasy.org/uniprot/P17080 RBMS1 RNA binding motif, single YC1; MSSP; SCR2; MSSP-1; 5937; stranded interacting MSSP-2; MSSP-3; http://ca.expasy.org/uniprot/P29558 protein 1, transcript MGC3331; MGC15146 variant RNF12 Ring finger protein 12, RLIM; MGC15161; NY-REN- 51132; transcript variant 1 43 http://ca.expasy.org/uniprot/Q9NVW2 RPL30 ribosomal protein L30 RPL30 6156; http://ca.expasy.org/uniprot/P04645 RPL31 ribosomal protein L31 MGC88191; RPL31 6160; http://ca.expasy.org/uniprot/P62899 RUFY1 RUN and FYVE domain RABIP4; ZFYVE12; 80230; containing 1 FLJ22251 http://ca.expasy.org/uniprot/Q96T51 SMN1 Survival of motor neuron SMA; SMN; SMA1; SMA2; 6606; 1, telomeric SMA3; SMA4; SMA@; http://ca.expasy.org/uniprot/Q16637 SMNT; BCD541; T-BCD541 SNK serum-inducible kinase SNK; PLK2 10769; http://ca.expasy.org/uniprot/Q9NYY3 SSNA1 Sjogren's syndrome N14; NA14; NA-14 8636; nuclear autoantigen 1 http://ca.expasy.org/uniprot/O43805 STK11 Serine/threonine kinase PJS; LKB1 6794; 11 (Peutz-Jeghers http://ca.expasy.org/uniprot/Q15831 syndrome) TOM1 Target of myb1 (chicken) FLJ33404 10043; http://ca.expasy.org/uniprot/O60784 TXNRD1 Thioredoxin reductase 1, TR; TR1; TXNR; TRXR1; 7296; transcript variant 5 GRIM-12; MGC9145 http://ca.expasy.org/uniprot/Q16881 VCL vinculin MVCL; CMD1W; VCL 7414; http://ca.expasy.org/uniprot/P18206

In other embodiments, a method is provided which is useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, or in the panel of auto-antigens comprises at least 10 auto-antigens selected from Table 3. In other embodiments, the at least 10 auto-antigens may be selected from Table 1, or from Table 2. Preferably a panel comprises at least 20 auto-antigens. Optionally a panel may comprise 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 or more auto-antigens.

The panel of auto-antigens useful in the diagnosis of lupus may comprise the auto-antigens set out in Table 1, or in Table 2, or in Table 3. In some embodiments the panel may have up to 5 auto-antigens deleted or replaced, or optionally up to 6, 7, 8, 9 or 10 or more auto-antigens deleted or replaced provided the required specificity and selectivity of the method is retained.

It is expected that a clinically valuable panel of biomarkers should be large enough to allow a disease to be detected accurately, minimizing false positives and false negatives, whilst also being small enough to be practical.

In embodiments the methods of the present invention use a panel of auto-antigens comprising the auto-antigens listed in Table 4. Such a method may use a panel comprising 1 or more additional auto-antigens selected from the group consisting of the auto-antigens of Table 5.

In the methods of the present invention detecting the presence or an increased level of at least 2 auto-antibodies provides information useful for diagnosis. Optionally, detecting the presence of an increased level of at least 3, preferably at least 4 and more preferably at least 5 auto-antibodies provides information useful for diagnosis.

By analysing the relationship between the panel of biomarkers and the status of the samples used to derive the panel, it is possible to derive a mathematical relationship between the presence, absence or a modulated level of auto-bodies directed against proteins in the panel and the presence, absence, progression or regression of lupus. The application of this mathematical relationship to naive samples to be tested can be used to derive information useful for diagnosis, progression or regression of lupus.

Where it is not possible to derive such a mathematical relationship, it is possible to derive a statistical relationship between the presence, absence or modulated level of auto-antibodies directed against proteins in the panel and the presence, absence, progression or regression of lupus. The application of this statistical relationship to naive sample to be tested can be used to derive information useful for diagnosis, progression or regression of lupus.

According to a different aspect of the present invention there is provided a diagnostic kit for use in diagnosing a patient as having or being predisposed to developing lupus, or having presymptomatic lupus, said kit comprising one or more auto-antigens selected from the group consisting of ZMAT2 (Zinc finger, matrin type 2); BANK1 (B-cell scaffold protein with ankyrin repeats 1); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); L1N28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11_(Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); or ZNF38 (Zinc finger protein 38), which one or more auto-antigens are immobilised on a substrate.

In one embodiment, the one or more auto-antigens includes ZMAT2.

In addition to one or more of the auto-antigens above, the kit may comprise one or more of the auto-antigens BANK1 (B-cell scaffold protein with ankyrin repeats 1); IFI16 (Interferon, gamma-inducible protein 16); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki), transc); RALBP1 (RalA-binding protein 1); and SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)).

Alternatively, said kit may comprise a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or more, even more preferably to a panel comprising all 35, auto-antigens selected from the auto-antigens listed in Table 1, wherein the auto-antigens are immobilised on a substrate.

According to another aspect of the present invention a diagnostic kit is provided for use in diagnosing a patient as having or being predisposed to developing lupus, or having pre-symptomatic lupus, said kit comprising one or more auto-antigens selected from the group of auto-antigens of Table 2.

Alternatively, said kit may comprise a panel comprising 5 or more, preferably 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 or more, even more preferably to a panel comprising all 32 auto-antigens selected from the auto-antigens listed in Table 2, wherein the auto-antigens are immobilised on a substrate.

According to another aspect of the present invention there is provided a diagnostic kit for use in diagnosing a patient as having or being predisposed to developing lupus, or having pre-symptomatic lupus, said kit comprising 1 or more auto-antigens selected from the group of auto-antigens of Table 3.

Alternatively said kit may comprise a panel comprising 5 or more, preferably, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or more, even more preferably to a panel comprising all 45 auto-antigens selected from the auto-antigens listed in Table 3, wherein the auto-antigens are immobilised on a substrate.

In some embodiments, said substrate may comprise a strip, a slide, beads or a well of a microtitre plate or a “chip” for use with Surface Plasmon Resonance (Biacore) or planar waveguide technology. Suitably, the substrate may comprise a disposable test strip of the kind widely used in diagnostic kits for use at the point of care.

The panel of auto-antigens may be carried on said substrate in the form of an addressable array. Said array may be a microarray. The auto-antigens of the panel may constitute at least 5%, 10%, 15%, 20%, 25%, 40% 50%, typically at least 70% or 80%, of the total number of proteins present on the array. For example, the auto-antigens of the panel may constitute more than 85%, 90% or 95% of the total number of proteins on the array.

Alternatively, said diagnostic kit may comprise said panel of auto-antigens carried on a substrate in the form of an array, which array includes one or more other proteins or polypeptides, and means for automatically analysing the results of tests performed using said array, said analysing means including computer-readable instructions in which results from said one or more other proteins or polypeptides are blinded during the analysis.

In addition to the auto-antigens (and optional one or more other proteins or polypeptides) forming said panel, the array may further comprise one or more control proteins or polypeptides. Suitably, the array may comprise one or more replicates of the auto-antigens or controls, preferably replicates of each auto-antigen and each control.

Advantageously, the array comprises an array of correctly folded auto-antigens, which may be prepared in accordance with the methods disclosed by WO-A-01/57198 or WO-A-02/27327, for example, the contents of which are incorporated herein by reference. Preferably, the array comprises an array of correctly folded tagged auto-antigens that are attached to said substrate via the tag, which array may be prepared, for example, in accordance with the methods disclosed by WO 03/064656 or WO-A-2004/046730, the contents of which are also incorporated herein by reference.

Said kit may further comprise an anti-human immunoglobulin antibody or other protein that specifically binds immunoglobulin, and means for detecting the anti-human immunoglobulin antibody or other immunoglobulin-binding protein.

Said detection means may comprise a colorimetric or fluorescent assay or utilise label-free methods such as surface plasmon resonance, mass spectrometry or planar waveguide technology.

The anti-human immunoglobulin antibody may be an anti-human IgM antibody, an anti-human IgG antibody, an anti-human IgA antibody, an anti-human IgE antibody or a combination of any two or more of such anti-human antibodies; preferably, the anti-human immunoglobulin antibody is an anti-human IgG antibody. Alternatively, or additionally, a protein that specifically binds an antibody is included in the kit. Preferably, this protein is protein A or protein G, or a recombinant form thereof.

According to yet another aspect of the present invention there is provided the use of one or more auto-antigens selected from the aforementioned groups as shown in Tables 1, 2, 3 or 4, or antigenic fragments or antigenic epitopes thereof, or a nucleic acid encoding the auto-antigen or fragment or epitope thereof, as an active agent in the manufacture of a vaccine for the treatment or prevention of lupus. Immunization with such an active agent may be protective against aberrant expression of the said antigen in lupus. In one embodiment the vaccine comprises ZMAT2, optionally in combination with one or more further antigens specified in Tables 1, 2, 3 or 4.

Said active agent may be formulated in a pharmaceutical composition in accordance with method is well known in the art. Said composition may further comprise a pharmaceutically acceptable vehicle. Said vehicle may comprise one or more excipients, carriers, buffers, stabilisers or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active agent. The precise nature of the carrier or other material may depend on the route of administration, e.g., oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal or patch routes.

Pharmaceutical compositions for oral administration may be in tablet, capsule, powder or liquid form. A tablet may include a solid carrier such as gelatin or an adjuvant. Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included.

For intravenous, cutaneous or subcutaneous injection, or injection at the site of affliction, the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability. Those of relevant skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection. Preservatives, stabilisers, buffers, antioxidants and/or other additives may be included, as required.

Whether it is a polypeptide, peptide, or nucleic acid molecule, other pharmaceutically useful compound according to the present invention that is to be given to an individual, administration is preferably in a “prophylactically effective amount” or a “therapeutically effective amount” (as the case may be, although prophylaxis may be considered therapy), this being sufficient to show benefit to the individual. The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of what is being treated. Prescription of treatment, e.g. decisions on dosage etc, is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disorder to be treated, the condition of the individual patient, the site of delivery, the method of administration and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in REMINGTON'S PHARMACEUTICAL SCIENCES, 16th edition, Osol, A. (ed), 1980.

Alternatively, targeting therapies may be used to deliver the active agent more specifically to certain types of cell, by the use of targeting systems such as antibody or cell specific ligands. Targeting may be desirable for a variety of reasons; for example if the agent is unacceptably toxic, or if it would otherwise require too high a dosage, or if it would not otherwise be able to enter the target cells.

Instead of administering these agents directly, they could be produced in the target cells by expression from an encoding gene introduced into the cells, e.g. in a viral vector (a variant of the VDEPT technique—see below). The vector could be targeted to the specific cells to be treated, or it could contain regulatory elements, which are switched on more or less selectively by the target cells.

Alternatively, the agent could be administered in a precursor form, for conversion to the active form by an activating agent produced in, or targeted to, the cells to be treated. This type of approach is sometimes known as ADEPT or VDEPT; the former involving targeting the activating agent to the cells by conjugation to a cell-specific antibody, while the latter involves producing the activating agent, e.g. a vaccine or fusion protein, in a vector by expression from encoding DNA in a viral vector (see for example, EP-A-415731 and WO 90/07936).

The vaccines of the present invention also include one or more adjuvant compounds, i.e. a compound or compounds that increases an immunogenic response or the immunogenicity of an antigen or vaccine. Adjuvant compounds useful in the present invention include Complete Freund's Adjuvant (CFA); Incomplete Freund's Adjuvant (IFA); Montanide ISA (incomplete seppic adjuvant); Ribi Adjuvant System (RAS); TiterMax; Syntex Adjuvant Formulation (SAF); Aluminum Salt Adjuvants; Nitrocellulose-adsorbed antigen; Encapsulated or entrapped antigens; Immune-stimulating complexes (ISCOMs); and Gerbu^(R) adjuvant.

Following is a description by way of example only with reference to the accompanying drawings of embodiments of the present invention.

In the drawings, FIG. 1 is a cartoon illustrating a method for the detection of auto-antibodies in serum.

EXAMPLES Example 1 A Microarray of Antigens for Discovering Auto-Antibody Biomarkers in Patient Serum that are Diagnostic for Lupus

Full-length open reading frames for 330 target genes covering a wide range of protein classes were cloned in-frame with a sequence encoding a C-terminal E. coli BCCP-myc tag (WO 03/064656; Boutell) in a baculovirus transfer vector and sequence-verified. Recombinant baculoviruses were generated, amplified and expressed in Sf9 cells grown in suspension using standard methods adapted for 24-well deep well plates (Whatman, Maidstone, UK; Chambers, Zhao). Recombinant protein expression was analyzed for protein integrity and biotinylation by Western blotting. Cells harboring recombinant protein were lysed and lysates were spotted in quadruplicate using a QArray2 Microarrayer (Genetix, New Milton, UK) equipped with 300 μm solid pins on to streptavidin-coated glass slides (Schott Nexterion, Jena, Germany). Slides were stored at −20° C. A range of protein activities have been demonstrated to be stable under these conditions for over 12 months. Proteins spotted on the array project into an aqueous environment and orient away from the surface of the glass chips, exposing them for binding by auto-antibodies.

In addition to the 330 proteins, four control proteins for the BCCP-myc tag (BCCP, BCCP-myc, β-galactosidase-BCCP-myc and β-galactosidase-BCCP) were arrayed, along with Cy3/Cy5-labeled biotin-BSA, dilution series of biotinylated-IgG and biotinylated IgM, a biotinylated-myc peptide dilution series and buffer only spots.

Example 2 Detection of Auto-Antibodies in Serum Samples from Patients Suffering from Lupus

Serum samples of 169 individuals diagnosed with lupus (median age 47 years, 95% females) were obtained from 2 commercial sources. As controls, samples of 170 individuals with no known history of lupus (median age 50 years, 95% females) were obtained. These samples were assayed for the presence of auto-antibodies using the protein arrays described in Example 1.

Serum samples were clarified (centrifuged at 10-13K rcf for 3 minutes at 4° C. to remove particulates, including lipids) and diluted 200-fold in 0.1% v/v Triton/0.1% w/v BSA in 1× PBS (Triton-BSA-PBS buffer) and then applied to the arrays. Each diluted serum sample (2.0 mL) was applied to a separate array and incubated in a dish for 2 hours at room temperature (RT, 20° C.) with gentle orbital shaking 50 rpm). Arrays were then carefully removed from the dish and any excess probing solution was removed by blotting the sides of the array onto lint-free tissue. Probed arrays were washed twice in fresh Triton-BSA-PBS buffer in a new plastic container with screw-on lid at room temperature for 5 minutes with rapid orbital shaking. The washed slides were then blotted onto lint-free tissue to remove excess wash buffer and were incubated in a secondary antibody solution (prepared just prior to use) at room temperature for 2 hours, with gentle orbital shaking and protected from light. The secondary antibody solution was a Cy3 labeled rabbit anti-human IgG antibody diluted in Triton-BSA-PBS buffer to give optimal detection of bound serum IgG. The slides were washed three times in Triton-BSA-PBS buffer for 5 minutes at RT with gentle orbital shaking, rinsed briefly (5-10 seconds) in distilled water, and centrifuged for 2 minutes at 240 rcf in a container suitable for centrifugation. To help wick away excess liquid on the arrays, a lint-free tissue was placed at the bottom of the arrays during centrifugation.

The probed and dried arrays were then scanned using a microarray scanner capable of using an excitation wavelength suitable for the detection of the secondary staining solution, such as the Molecular Devices Genepix 4000B microarray scanner, to identify auto-antibodies bound by the array and to determine intensity of auto-antibody binding. The microarray scans produced TIFF images for each array that were used to determine the intensity of fluorescence bound to each protein spot.

Raw median signal intensity (also referred to as the relative fluorescent unit, RFU) of each protein feature (also referred to as a spot or antigen) on the array was determined from the TIFF images using GenePix Pro microarray data analysis software or similar software and subtracted from the local median background intensity. It is also possible to use other measures of spot intensity such as the mean fluorescence, total fluorescence, as is known by anyone skilled in the art.

The resulting net fluorescent intensities of all protein features on each array were then normalized using the quantile normalization (Bolstad) method. Data for sera from two lupus patients and one control serum were excluded because many of their antigen binding signals were masked by high background binding. Other methods for data normalization suitable for the current data include, amongst others, multiplication of net fluorescent intensities by a normalization factor consisting of the product of the 1^(st) quartile of all intensities of a sample and the mean of the 1^(st) quartiles of all samples and the vsn method (Huber). Such normalization methods are known to anyone skilled in the art of microarray analysis. The normalized fluorescent intensities were then averaged for each protein feature.

The normalized mean data were used to select biomarkers for lupus. The data were randomly split into training and test sets 10 times (10 fold cross-validation)

Proteins were selected and ranked with the significance analysis of microarrays (SAM) algorithm (Tusher) using the data in the training subset. Similar methods that are applicable to this task include, amongst others, Prediction Analysis of Microarrays (PAM, Tibshirani), MP test (Fox), fold change (Witten) and principal component analysis (Bessant). Such data analysis methods are known to anyone skilled in the art of microarray analysis. The data were analysed as two class—unpaired (diseased vs control) using a non-parametric test statistic. The protein features selected in each cross-validation fold with the highest scores and a q-value of ≦30% were selected and those with q-values≦30% in most of the 10 cross-validation iterations were selected and are shown in Table 2 alongside the frequency of selection in cross-validation.

Table 1a lists the protein features for which the q-value was ≦30% by SAM analysis alongside the selection frequency in cross-validation

Gene symbol cross-validation selection frequency ZNF38 10 BPY2IP1 10 CEBPG 10 E1B-AP5 10 FUS 10 HMG20B 10 LIN28 10 NDUFV3 10 PABPC1 10 PHLDA1 10 PIAS2 10 PSME3 10 RAB11FIP3 10 RALBP1 10 RAN 10 RARA 10 RBMS1 10 RNF12 10 SSA2 10 STAU 10 STK11 10 TOM1 10 ZMAT2 10 SRPK1 9 TXNRD1 9 BANK1 8 IFI16 8 KRT8 8 RUFY1 8 SMN1 8 SSNA1 8 HAGH 7 HOXB6 7 RDBP 7 TXNL2 7

The proteins listed in Table la are more frequently bound by sera from diseased than control subjects. The proteins listed in Table la are considered as biomarkers useful in diagnosis of lupus.

The normalized data of the training set for the proteins selected in each cross-validation fold were used to construct a classification algorithm. An algorithm was established based on using logistic regression (Liao) algorithms. Similar methods that can be used to construct classification algorithms for the sample data of this study include, amongst others, PAM (Tibshirani), artificial neural networks (Bessant), support vector machine (SVM) (Brown) and regularized discriminant analysis (Guo). Such data analysis methods are known to anyone skilled in the art of microarray analysis. The constructed algorithm was then used to classify the samples of the test set into diseased and control samples in order to check its validity as classification method for lupus. In order to get an estimate of the classification errors the mean specificity and sensitivity were examined using 10 fold cross validation. The cross validation mean sensitivity was 67%, the mean specificity was 85%. The cross validation approach ensured robust measurements of classification errors.

Example 3 Detection of Auto-Antibodies in Serum Samples from Patients Suffering from Lupus Using a Modified Analysis Approach

The raw data (169 lupus and 170 controls samples) as described in Example 2 was subjected to modified analysis in order to mine the data still further and identify additional markers indicative of lupus.

Raw median signal intensity (relative fluorescent unit, RFU) of each protein feature on the array was determined from the TIFF images using ArrayPro microarray data analysis software and subtracted from the local median background intensity.

The resulting net fluorescent intensities of all protein features from all three array types were then normalized together using the vsn normalization method (Huber 2002), or multiplication of net fluorescent intensities by a normalization factor consisting of the product of the 1st quartile of all intensities of a sample and the mean of the 1st quartiles of all samples, or the quantile or robust quantile method (Bolstad 2003). The normalized fluorescent intensities were then averaged for each protein feature.

The normalized mean data were used to select biomarkers for lupus. In order to estimate the specificity and sensitivity of a classification method based on biomarkers selected, data were initially split randomly into training and test sets 10 times (10 fold cross-validation).

Proteins were selected and ranked with the MP test (Fox 2006), or Prediction Analysis of Microarrays (PAM, Tibshirani 2002), significance analysis of microarrays (SAM) algorithm (Tusher 2001, Hueber 2005), fold change (Witten 2007), or a cut-off method based on a confidence interval derived from the data of the controls, using the data in the training subset. The data were analysed as two class (diseased vs control).

The normalized data of the training set for the proteins selected in each cross-validation fold were used to construct a classification algorithm. Features derived from the same clones that were selected on more than one array type were left in for the classification algorithm construction. An algorithm was established based on penalised logistic regression (Antoniadis 2003, Liao 2007), or PAM (Tibshirani 2002), or support vector machine (SVM, Brown 2000, Statnikov 2008) or a cut-off method based on a confidence interval derived from the data of the controls. The constructed algorithms were then used to classify the samples of the test set into diseased and control samples in order to check its validity as classification method for lupus. In order to get an estimate of the classification errors the mean specificity and sensitivity were examined using 10 fold cross validation.

The proteins selected by SAM analysis in at least 9 of the 10 cross validation folds are shown in Table 2a. The analysis parameters used were: q-value cut-off ≦20% and proteins with elevated responses included.

Table 2a lists the protein features for which the q-value was ≦20% by SAM analysis and 10 fold cross validation selection frequency ≧9.

Gene symbol Selection frequency ASPSCR1 10 BANK1 10 CEBPG 10 DDX55 10 DOM3Z 9 E1B-AP5 10 FUS 10 HMG20B 10 IFI16 10 KRT8 10 LIN28 10 LNX 10 MAP1S 10 PABPC1 10 PHLDA1 10 PIAS2 10 PRKCBP1 10 PRKRA 10 PSME3 10 RALBP1 10 RARA 10 RDBP 10 RPL31 9 RPL30 10 SNK 10 SRPK1 10 SSA2 10 STAU1 10 TXNL2 10 VCL 10 ZMAT2 9 ZNF38 9

The proteins listed in Table 2a are more frequently bound by sera from diseased than control subjects. The proteins listed in Table 2a are considered as biomarkers useful in diagnosis of lupus.

Classification by SVM gave a cross validation mean sensitivity of 74% and a mean specificity of 86%.

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Gene Symbol, Entrez Gene ID, DNA Accession Number and Sequence of Antigens.

Gene symbol: ASPSCR1 Entrez Gene ID: 79058 dna/ac: BC018722.1 sequence: (SEQ ID NO: 1) atggcggccccggcaggcggcggaggctccgcggtgtcggtgctggcc ccgaacggccggcgccacacggtgaaggtgacgccgagcaccgtgctg cttcaggttctggaggacacgtgccggcggcaggacttcaacccctgt gaatatgatctgaagtttcagaggagcgtgctcgacctttctctccag tggagatttgccaacctgcccaacaatgccaagctggagatggtgccc gcttcccggagccgtgaggggcctgagaacatggttcgcatcgctttg cagctggacgatggctcgaggttgcaggactattctgttcaggccaga ccctctgggagatctcagccattttccacagatcagggagtgcctgca gcaccccggcggggccaccccagtctgcgtgtacacgagggatgaggt gacgggtgaagctgccctgcggggcacgacgctgcagtcgctgggcct gaccgggggcagcgccaccatcaggtttgtcatgaagtgctacgaccc cgtgggcaagaccccaggaagcctgggctcgtcagcgtcggctggcca ggcagccgccagcgctccacttccatggaatctggggagctcagccgc ggcgacttgagccgtccggaggacgcggacacctcagggccctgctgc gagcacactcaggagaagcagagcacaagggcacccgcagctgcccca ttgttcattctcgggtgggggacagagacaggggggccctcctgggcc cacgaggcctctgacatcatcttcagctaagttgccgaagtccctctc cagccctggaggcccctccaagccaaagaagtccaagtcgggccagga tccccagcaggagcaggagcaggagcgggagcgggatccccagcagga gcaggagcgggagcggcccgtggaccgggagcccgtggaccgggagcc ggtggtgtgccaccccgacctggaggagcggctgcaggcctggccagc ggagctgcctgatgagttattgagctgacggtggacgacgtgagaaga cgcttggcccagctcaagagtgagcggaagcgcctggaagaagccccc ttggtgaccaaggccttcagggaggcgcagataaaggagaagctggag cgctacccaaaggtggctctgagggtcctgttccccgaccgctacgtc ctacagggcttatccgccccagcgagacagtgggggacttgcgagact tcgtgaggagccacctggggaaccccgagctgtcattttacctgttca tcacccctccaaaaacagtcctggacgaccacacgcagaccctctttc aggcgaacctcttcccggccgctctggtgcacttgggagccgaggagc cggcaggtgtctacctggagcctggcctgctggagcatgccatctccc catctgcggccgacgtgctggtggccaggtacatgtccagggccgccg ggtccccttccccattgccagcccctgaccctgcacctaagtctgagc cagctgctgaggagggggcgctggtcccccctgagcccatcccaggga cggcccagcccgtgaagaggagcctgggcaaggtgcccaagtggctga agctgccggccagcaagaggtga Gene symbol: BANK1 Entrez Gene ID: 55024 dna/ac: BC032241.1 sequence: (SEQ ID NO: 2) atgctgccagcagcgccaggcaaggggcttgggagcccggaccccgcc ccctgcggcccagcgcccccaggaaatacaaaagatataataatgata tatgaagaagatgctgaggaatgggctctgtacttgacagaagtattt ttacatgttgtgaaaagggaagccatcctgttatatcgcttggagaat ttctcttttcggcatttggagttgctgaacttaacgtatacaaatgta aacttttgatattatcaaatagcctgcttagagacctaactccaaaga aatgtcagtttctggaaaagatacttcattcaccaaaaagtgtagtta ctttgctttgtggagtgaagagttcagatcagctctatgaattactaa atatctctcaaagcagatgggagatctcaactgaacaggaacctgaag actacatctctgtaatccagagtatcatattcaaagattctgaagact actttgaggtcaacattccaacagacctacgagcaaaacattctgggg aaataagtgagagaaaggaaattgaagaactatcagaagatcaagaaa caccataccactagcagtggtgcttcccactgaaattccatgtgagaa tcctggtgaaatattcataattttgagagatgaagtaattggtgatac tgtagaggttgaatttacatcaagtaataagcgcattagaacacggcc agccattggaataagaaagtctggtgcatgaaagctttagagtttcct gctggttcagtccatgtcaatgtctactgtgatggaatcgttaaagct acaaccaaaattaagtactacccaacagcaaaggcaaaggaatgccta ttcagaatggcagattcaggagagagtttgtgccagaatagcattgaa gaacttgatggtgtccttacatccatattcaaacatgagataccatat tatgagttccagtctcttcaaactgaaatttgttctcaaaacaaatat actcatttcaaagaacttccaactcttctccactgtgcagcaaaattt ggcttaaagaacctggctattcatttgcttcaatgttcaggagcaacc tgggcatctaagatgaaaaatatggagggttcagaccccgcacatatt gctgaaaggcatggtcacaaagaactcaagaaaatcttcgaagacttt tcaatccaagaaattgacataaataatgagcaagaaaatgattatgaa gaggatattgcctcattttccacatatattccttccacacagaaccca gcatttcatcatgaaagcagaaagacatacgggcagagtgcagatgga gctgaggcaaatgaaatggaaggggaaggaaaacagaatggatcaggc atggagaccaaacacagcccactagaggttggcagtgagagttctgaa gaccagtatgatgacttgtatgtgttcattcctggtgctgatccagaa aataattcacaagagccactcatgagcagcagacctcctctccccccg ccgcgacctgtagctaatgccttccaactggaaagacctcacttcacc ttaccagggacaatggtggaaggccaaatggaaagaagtcaaaactgg ggtcatcctggtgttagacaagaaacaggagatgaacccaaaggagaa aaagagaagaaagaagaggaaaaagagcaggaggaggaagaagaccca tatacttttgctgagattgatgacagtgaatatgacatgatattggcc aatctgagtataaagaaaaaaactgggagtcggtctttcattataaat agacctcctgcccccacaccccgacccacaagtatacctccaaaagag gaaactacaccttacatagctcaagtgtttcaacaaaagacagccaga agacaatctgatgatgacaagttccgtggtcttcctaagaaacaagac agagctcggatagagagtccagccttttctactctcaggggctgtcta actgatggtcaggaagaactcatcctcctgcaggagaaagtaaagaat gggaaaatgtctatggatgaagctctggagaaatttaaacactggcag atgggaaaaagtggcctggaaatgattcagcaggagaaattacgacaa ctacgagactgcattattgggaaaaggccagaagaagaaaatgtctat aataaactcaccattgtgcaccatccaggtggtaaggaaactgcccac aatgaaaataagtttttataatgtacacttcagcaataagcttcctgc tcgaccccaagttgaaaaggaatttggtttctgttgcaagaaagatca ttaa Gene symbol: CEBPG Entrez Gene ID: 1054 dna/ac: BC007582.1 sequence: (SEQ ID NO: 3) atgagcaagatatcgcagcaaaacagcactccaggggtgaacggaatt agtgttatccatacccaggcacatgccagcggcttacagcaggttcct cagctggtgcctgctggccctgggggaggaggcaaagctgtggctccc agcaagcagagcaaaaagagttcgcccatggatcgaaacagtgacgag tatcggcaacgccgagagaggaacaacatggctgtgaaaaagagccgg ttgaaaagcaagcagaaagcacaagacacactgcagagagtcaatcag ctcaaagaagagaatgaacggttggaagcaaaaatcaaattgctgacc aaggaattaagtgtactcaaagatttgtttcttgagcatgcacacaac cttgcagacaacgtacagtccattagcactgaaaatacgacagcagat ggcgacaatgcaggacagtag Gene symbol: DDX55 Entrez Gene ID: 57696 dna/ac: BC030020.2 sequence: (SEQ ID NO: 4) atggagcatgtgacagagggctcctgggagtcgctgcctgtgccgctg cacccgcaggtgctgggcgcgctgcgggagctgggcttcccgtacatg acgccggtgcagtccgcaaccatccctctgttcatgcgaaacaaagat gtcgctgcagaagcggtcacaggtagtggcaaaacactcgcttttgtc atccccatcctggaaattcttctgagaagagaagagaagttaaaaaag agtcaggttggagccataatcatcacccccactcgagagctggccatt caaatagacgaggtcctgtcgcatttcacgaagcacttccccgagttc agccagattctttggatcggaggcaggaatcctggagaagatgttgag aggtttaagcaacaaggtgggaacatcattgtggccactccaggccgc ttggaggacttgttccggaggaaggccgaaggcttggatctggccagc tgtgtgcgatccctggatgtcctggtgttggatgaggcagacagactt ctggacatggggtttgaggcaagcatcaacaccattctggagtttttg ccaaagcagaggagaacaggccttttctctgccactcagacgcaggaa gtggagaacctggtgagagcgggcctccggaaccctgtccgggtctca gtgaaggagaagggcgtggcagccagcagtgcccagaagaccccctcc cgcctggaaaactactacatggtatgcaaggcagatgagaaatttaat cagctggtccattttcttcgcaatcataagcaggagaaacacctggtc ttcttcagcacctgtgcctgtgtggaatactatgggaagactctggaa gtgctggtgaagggcgtgaagattatgtgcattcacggaaagatgaaa tataaacgcaataagatcttcatggagttccgcaaattgcaaagtggg attttagtgtgcactgatgtgatggcccggggaattgatattcctgaa gtcaactgggttttgcagtatgaccctcccagcaatgcaagtgccttc gtgcatcgctgcggtcgcacagctcgcattggccacgggggcagcgct ctggtgttcctectgcccatggaagagtcatacatcaatttccttgca attaaccaaaaatgccccctgcaggagatgaagccccagagaaacaca gcggaccttctgccaaaactcaagtccatggccctggctgacagagag tgtttgaaaagggcatgaaagcttttgtgtcatatgtccaagcttatg caaagcatgaatgcaacctgattttcagattaaaggatcttgattttg ccagccttgctcgaggttttgccctgctgaggatgcccaagatgccag aattgagagggaagcagtttccagattttgtgcccgtggacgttaata ccgacacgattccatttaaagataaaatcagagaaaagcagaggcaga aactcctggagcaacaaagaagagagaaaacagaaaatgaagggagaa gaaaattcataaaaaataaagcttggtcaaagcagaaggccaaaaaag aaaagaagaaaaaaatgaatgagaaaaggaaaagggaagagggttctg atattgaagatgaggacatggaagaacttcttaatgacacaagactct tgaaaaaacttaagaaaggcaaaataactgaagaagaatttgagaagg gcttgttgacaactggcaaaagaacaatcaagacagtggatttaggga tctcagatttggaagatggctgctaa Gene symbol: DOM3Z Entrez Gene ID: 1797 dna/ac: BC019083.2 sequence: (SEQ ID NO: 5) atggatcccagggggaccaagagaggagctgagaagacagaggtagct gagcctcggaacaaactacctcgtccagcaccttctctgcccacagac cctgccctotactctgggccctttcattctaccggcgcccttcggaac tgggctgcttctccctggatgctcaacgccagtaccatggagatgccc gagccctgcgctactatagcccaccccccactaacggtccaggcccca actttgacctcagagacggatacccggatcgataccagccccgggacg aggaggtccaggaaaggctggaccacctgctgtgctggctcctggaac accgaggccggttggaggggggtccaggctggctggcagaggccatag tgacgtggcgggggcacctgacaaaactgctgacgacaccgtatgagc ggcaggagggctggcagctggcagcctcccggttccagggaacactat acctgagtgaagtggagacaccgaacgctcgggcccagaggcttgacg gccaccgctcctccgggagcttatgtacatgggatacaaatttgagca gtacatgtgtgcagacaaacctggaagctccccagacccctctgggga ggttaacaccaacgtggccttctgactgtgctacgcagccgcctggga agccaccactgctcttctcaggggaggtagactgcacagacccccaag ccccatccacacagcccccaacctgctatgtggagctcaagacctcca aggagatgcacagccctggccaatggaggagtactacagacacaagct cctgaaatggtgggctcagtcattcctcccaggggtcccgaatgttgt tgctggcttccgtaacccagacggttttgtacttccctcaagaccttt cctaccatgaagatgtttgaatatgtcaggaatgaccgtgacggctgg aatccctctgtgtgcatgaacttctgtgccgccttccttagctttgcc cagagcacggttgtccaggatgaccccaggctcgttcatctcttctct tgggagcctggcggcccagtcaccgtgtctgtacaccaagatgcacct tacgccttcctgcccatatggtatgtggaagctatgactcaggacctc ccatcaccccccaagactccctctcccaaatag Gene symbol: E1B-AP5 Entrez Gene ID: 11100 dna/ac: BC009988.2 sequence: (SEQ ID NO: 6) atggacaatattaccaggcagaaccaattctacgatacccaagtcatc aaacaagaaaacgagtcaggctacgagaggagaccactggaaatggag cagcagcaggcctatcgtccagaaatgaagacagagatgaagcaagga gcacccaccagcttcctcccgcctgaagcttctcaactcaagccagac aggcagcaattccagagtcgaaagaggccttatgaagaaaaccgggga cgggggtactttgagcaccgagaggataggaggggccgctctcctcag cctcctgctgaagaggatgaagatgactttgatgatacccttgttgct attgacacctataactgcgacctccacttcaaggtggcccgagatcgg agtagtggctatccgctcacaattgagggctttgcatacctgtggtca ggagcccgtgccagctatggggtcagaaggggccgtgtatgcttcgag atgaagatcaatgaggaaatctccgtgaagcaccttccgtctacagag cctgacccccacgtggtccgtatcggctggtccctggactcctgcagc acccagctaggcgaagagcctttctcctatggctatggaggcactggg aagaagtccaccaatagccggtttgaaaactacggagacaagtttgca gagaacgatgtgattggctgctttgcggattttgaatgtggaaatgac gtggaactgtcttttaccaagaatggaaagtggatgggcattgctttc cgaatccagaaggaagccttggggggtcaggccctctatcctcatgtc ctggtgaagaattgcgcagtggagttcaacttcggacagagagcagag ccctactgttctgtcctcccggggtttaccttcatccagcaccttccc cttagtgagcgtatccggggcaccgttggaccaaagagcaaggcagaa tgtgagattctgatgatggtgggcctgcctgctgctggcaagaccaca tgggccatcaaacatgcagcctccaacccttccaagaagtacaacatc ctgggtaccaatgccatcatggataagatgcgggtgatgggcctacgc cggcagcggaactatgctggccgctgggatgtcctgatccagcaggcc acccagtgcctcaaccgcctcatccagattgctgcccgcaagaaacgc aactatatcctagatcagacaaatgtttatgggtcagcccagagacga aaaatgagaccatttgaaggcttccagcgcaaagctattgtaatttgt cccactgacgaggacctaaaagaccgaacaataaagcgaaccgacgag gaagggaaggatgtcccagatcatgcggtcttagaaatgaaagccaac ttcacgttgccagatgttggggacttcctggatgaggttctgttcatt gagctgcagcgggaggaagcggacaagctagtgaggcagtacaacgag gaaggccgcaaggctgggccaccccctgaaaagcgctttgacaaccga ggtggtggtggcttccggggccgcgggggtggtggtggcttccagcgc tatgaaaaccgaggaccccctggaggcaaccgtggcggcttccagaac cgagggggaggcagcggtggaggaggcaactaccgaggaggtttcaac cgcagcggaggtggtggctatagccagaaccgctggggtaacaacaac cgggataacaacaactccaacaacagaggcagctacaaccgggctccc cagcaacagccgccaccacagcagcctccgccaccacagccaccaccc cagcagccaccgccaccacccagctacagccctgctcggaacccccca ggggccagcacctacaataagaacagcaacatccctggctcaagcgcc aataccagcacccccaccgtcagcagctacagccctccacagccgagt tacagccagccaccctacaaccagggaggttacagccagggctacaca gccccaccgcctccacctccaccaccacctgcctacaactatgggagc tacggcggttacaacccggccccctataccccaccgccaccccccacc gcacagacctaccctcagcccagctataaccagtatcagcagtatgcc cagcagtggaaccagtactatcagaaccagggccagtggccgccatac tacgggaactacgactacgggagctactccgggaacacacagggtggc acaagtacacagtag Gene symbol: FUS Entrez Gene ID: 2521 dna/ac: BC000402.2 sequence: (SEQ ID NO: 7) atggcctcaaacgattatacccaacaagcaacccaaagctatggggcc taccccacccagcccgggcagggctattcccagcagagcagtcagccc tacggacagcagagttacagtggttatagccagtccacggacacttca ggctatggccagagcagctattcttcttatggccagagccagaacaca ggctatggaactcagtcaactccccagggatatggctcgactggcggc tatggcagtagccagagctcccaatcgtcttacgggcagcagtcctcc tatcctggctatggccagcagccagctcccagcagcacctcgggaagt tacggtagcagttctcagagcagcagctatgggcagccccagagtggg agctacagccagcagcctagctatggtggacagcagcaaagctatgga cagcagcaaagctataatccccctcagggctatggacagcagaaccag tacaacagcagcagtggtggtggaggtggaggtggaggtggaggtaac tatggccaagatcaatcctccatgagtagtggtggtggcagtggtggc ggttatggcaatcaagaccagagtggtggaggtggcagcggtggctat ggacagcaggaccgtggaggccgcggcaggggtggcagtggtggcggc ggcggcggcggcggtggtggttacaaccgcagcagtggtggctatgaa cccagaggtcgtggaggtggccgtggaggcagaggtggcatgggcgga agtgaccgtggtggcttcaataaatttggtggccctcgggaccaagga tcacgtcatgactccgaacaggataattcagacaacaacaccatcttt gtgcaaggcctgggtgagaatgttacaattgagtctgtggctgattac ttcaagcagattggtattattaagacaaacaagaaaacgggacagccc atgattaatttgtacacagacagggaaactggcaagctgaagggagag gcaacggtctcttttgatgacccaccttcagctaaagcagctattgac tggtttgatggtaaagaattctccggaaatcctatcaaggtctcatgg ctactcgccgggcagactttaatcggggtggtggcaatggtcgtggag gccgagggcgaggaggacccatgggccgtggaggctatggaggtggtg gcagtggtggtggtggccgaggaggatttcccagtggaggtggtggcg gtggaggacagcagcgagctggtgactggaagtgtcctaatcccacct gtgagaatatgaacttctcttggaggaatgaatgcaaccagtgtaagg cccctaaaccagatggcccaggagggggaccaggtggctctcacatgg ggggtaactacggggatgatcgtcgtggtggcagaggaggctatgatc gaggcggctaccggggccgcggcggggaccgtggaggcttccgagggg gccggggtggtggggacagaggtggattggccctggcaagatggattc caggggtgagcacagacaggatcgcagggagaggccgtattaa Gene symbol: HAGH Entrez Gene ID: 3029 dna/ac: BC000840.1 sequence: (SEQ ID NO: 8) atgaaggtagaggtgctgcctgccctgaccgacaactacatgtacctg gtcattgatgatgagaccaaggaggctgccattgtggatccggtgcag ccccagaaggtcgtggacgcggcgagaaagcacggggtgaaactgacc acagtgctcaccacccaccaccactgggaccatgctggcgggaatgag aaactggtcaagctggagtcgggactgaaggtgtacgggggtgacgac cgtatcggggccctgactcacaagatcactcacctgtctacactgcag gtggggtctctgaacgtcaagtgcctggcgaccccgtgccacacttca ggacacatttgttacttcgtgagcaagcccggaggctcggagccccct gccgtgttcacaggtgacaccttgtttgtggctggctgcgggaagttc tatgaagggactgcggatgagatgtgtaaagctctgctggaggtcttg ggccggctccccccggacacaagagtctactgtggccacgagtacacc atcaacaacctcaagtttgcacgccacgtggagcccggcaatgccgcc atccgggagaagctggcctgggccaaggagaagtacagcatcggggag cccacagtgccatccaccctggcagaggagtttacctacaacccatca tgagagtgagggagaagacggtgcagcagcacgcaggtgagacggacc cggtgaccaccatgcgggccgtgcgcagggagaaggaccagttcaaga tgccccgggactga Gene symbol: HMG20B Entrez Gene ID: 10362 dna/ac: BC002552.2 sequence:  (SEQ ID NO: 9) atgtcccacggccccaagcagcccggcgcggccgccgcgccggcgggc ggcaaggctccgggccagcatgggggcttcgtggtgactgtcaagcaa gagcgcggcgagggtccacgcgcgggcgagaaggggtcccacgaggag gagccggtgaagaaacgcggctggcccaagggcaagaagcggaagaag attctgccgaatgggcccaaggcaccggtcacgggctacgtgcgcttc ctgaacgagcggcgcgagcagatccgcacgcgccacccggatctgccc tttcccgagatcaccaagatgctgggcgccgagtggagcaagctgcag ccaacggaaaagcagcggtacctggatgaggccgagagagagaagcag cagtacatgaaggagctgcgggcgtaccagcagtctgaagcctataag atgtgcacggagaagatccaggagaagaagatcaagaaagaagactcg agctctgggctcatgaacactctcctgaatggacacaagggtggggac tgcgatggcttctccaccttcgatgttcccatcttcactgaagagtta tggaccaaaacaaagcgcgtgaggeggagcttcggcgcttgcggaaga tgaatgtggccttcgaggagcagaacgcggtactgcagaggcacacgc agagcatgagcagcgcgcgcgagcgtctggagcaggagctggcgctgg aggagcggaggacgctggcgctgcagcagcagctccaggccgtgcgcc aggcgctcaccgccagettcgcctcactgccggtgccgggcacgggcg aaacgcccacgctgggcactctggacttctacatggcccggatcacgg agccatcgagcgcgaccccgcccagcacgagaagctcatcgtccgcat caaggaaatcctggcccaggtcgccagcgagcacctgtga Gene symbol: HOXB6 Entrez Gene ID: 3216 dna/ac: BC014651.1 sequence: (SEQ ID NO: 10) atgagttcctatttcgtgaactccaccttccccgtcactctggccagc gggcaggagtccttcctgggccagctaccgctctattcgtcgggctat gcggacccgctgagacattaccccgcgccctacgggccagggccgggc caggacaagggctttgccacttcctcctattacccgccggcgggcggt ggctacggccgagcggcgccctgcgactacgggccggcgccggccttc taccgcgagaaagagtcggcctgcgcactctccggcgccgacgagcag cccccgttccaccccgagccgcggaagtcggactgcgcgcaggacaag agcgtgttcggcgagacagaagagcagaagtgctccactccggtctac ccgtggatgcagcggatgaattcgtgcaacagttcctcctttgggccc agcggccggcgaggccgccagacatacacacgttaccagacgctggag ctggagaaggagtttcactacaatcgctacctgacgcggcggcggcgc atcgagatcgcgcacgccctgtgcctgacggagaggcagatcaagata tggttccagaaccgacgcatgaagtggaaaaaggagagcaaactgctc agcgcgtctcagctcagtgccgaggaggaggaagaaaaacaggccgag tga Gene symbol: IFI16 Entrez Gene ID: 3428 dna/ac: BC017059.1 sequence: (SEQ ID NO: 11) atgggaaaaaaatacaagaacattgttctactaaaaggattagaggtc atcaatgattatcatttttagaatggttaagtccttactgagcaacga tttaaaacttaatttaaaaatgagagaagagtatgacaaaattcagat tgctgacttgatggaagaaaagttccgaggtgatgctggtttgggcaa actaataaaaattttcgaagatataccaacgcttgaagacctggctga aactcttaaaaaagaaaagttaaaagtaaaaggaccagccctatcaag aaagaggaagaaggaagtggatgctacttcacctgcaccctccacaag cagcactgtcaaaactgaaggagcagaggcaactcctggagctcagaa aagaaaaaaatcaaccaaagaaaaggctggacccaaagggagtaaggt gtccgaggaacagactcagcctccctctcctgcaggagccggcatgtc cacagccatgggccgttccccatctcccaagacctcattgtcagctcc acccaacacttcttcaactgagaacccgaaaacagtggccaaatgtca ggtaactcccagaagaaatgttctccaaaaacgcccagtgatagtgaa ggtactgagtacaacaaagccatttgaatatgagaccccagaaatgga gaaaaaaataatgtttcatgctacagtggctacacagacacagttctt ccatgtgaaggttttaaacaccagcttgaaggagaaattcaatggaaa gaaaatcatcatcatatcagattatttggaatatgatagtctcctaga ggtcaatgaagaatctactgtatctgaagctggtcctaaccaaacgtt tgaggttccaaataaaatcatcaacagagcaaaggaaactctgaagat tgatattcttcacaaacaagcttcaggaaatattgtatatggggtatt tatgctacataagaaaacagtaaatcagaagaccacaatctacgaaat tcaggatgatagaggaaaaatggatgtagtggggacaggacaatgtca caatatcccctgtgaagaaggagataagctccaacttttctgctttcg acttagaaaaaagaaccagatgtcaaaactgatttcagaaatgcatag ttttatccagataaagaaaaaaacaaacccgagaaacaatgaccccaa gagcatgaagctaccccaggaacagagtcagcttccaaatccttcaga ggccagcacaaccttccctgagagccatcttcggactcctcagatgcc accaacaactccatccagcagtttcttcaccaagaaaagtgaagacac aatctccaaaatgaatgacttcatgaggatgcagatactgaaggaagg gagtcattttccaggaccgttcatgaccagcataggcccagctgagag ccatccccacactcctcagatgcctccatcaacaccaagcagcagttt cttaaccacgttgaaaccaagactgaagactgaacctgaagaagtttc catagaagacagtgcccagagtgacctcaaagaagtgatggtgctgaa cgcaacagaatcatttgtatatgagcccaaagagcagaagaaaatgtt tcatgccacagtggcaactgagaatgaagtcttccgagtgaaggtttt taatattgacctaaaggagaagttcaccccaaagaagatcattgccat agcaaattatgtttgccgcaatgggttcctggaggtatatcctttcac acttgtggctgatgtgaatgctgaccgaaacatggagatcccaaaagg attgattagaagtgccagcgtaactcctaaaatcaatcagctttgctc acaaactaaaggaagttttgtgaatggggtgtttgaggtacataagaa aaatgtaaggggtgaattcacttattatgaaatacaagataatacagg gaagatggaagtggtggtgcatggacgactgaccacaatcaactgtga ggaaggagataaactgaaactcacctgctttgaattggcaccgaaaag tgggaataccggggagttgagatctgtaattcatagtcacatcaaggt catcaagaccaggaaaaacaagaaagacatactcaatcctgattcaag tatggaaacttcaccagactttttcttctaa Gene symbol: KRT8 Entrez Gene ID: 3856 dna/ac: BC008200.1 sequence: (SEQ ID NO: 12) atgaacaaggtagagctggagtctcgcctggaagggctgaccgacgag atcaacttcctcaggcagctatatgaagaggagatccgggagctgcag tcccagatctcggacacatctgtggtgctgtccatggacaacagccgc tccctggacatggacagcatcattgctgaggtcaaggcacagtacgag gatattgccaaccgcagccgggctgaggctgagagcatgtaccagatc aagtatgaggagctgcagagcctggctgggaagcacggggatgacctg cggcgcacaaagactgagatctctgagatgaaccggaacatcagccgg ctccaggctgagattgagggcctcaaaggccagagggcttccctggag gccgccattgcagatgccgagcagcgtggagagctggccattaaggat gccaacgccaagttgtccgagctggaggccgccctgcagcgggccaag caggacatggcgcggcagctgcgtgagtaccaggagctgatgaacgtc aagctggccctggacatcgagatcgccacctacaggaagctgctggag ggcgaggagagccggctggagtctgggatgcagaacatgagtattcat acgaagaccaccagcggctatgcaggtggtctgagctcggcctatggg ggcctcacaagccccggcctcagctacagcctgggctccagattggct ctggcgcgggctccagctccttcagccgcaccagctcctccagggccg tggttgtgaagaagatcgagacacgtgatgggaagctggtgtctgagt cctctgacgtcctgcccaagtga Gene symbol: LIN28 Entrez Gene ID: 79727 dna/ac: BCO28566.2 sequence: (SEQ ID NO: 13) atgggctccgtgtccaaccagcagtttgcaggtggctgcgccaaggcg gcagaagaggcgcccgaggaggcgccggaggacgcggcccgggcggcg gacgagcctcagctgctgcacggtgcgggcatctgtaagtggttcaac gtgcgcatggggttcggcttcctgtccatgaccgcccgcgccggggtc gcgctcgaccccccagtggatgtctttgtgcaccagagtaagctgcac atggaagggttccggagcttgaaggagggtgaggcagtggagttcacc tttaagaagtcagccaagggtctggaatccatccgtgtcaccggacct ggtggagtattctgtattgggagtgagaggcggccaaaaggaaagagc atgcagaagcgcagatcaaaaggagacaggtgctacaactgtggaggt ctagatcatcatgccaaggaatgcaagctgccaccccagcccaagaag tgccacttctgccagagcatcagccatatggtagcctcatgtccgctg aaggcccagcagggccctagtgcacagggaaagccaacctactttcga gaggaagaagaagaaatccacagccctaccctgctcccggaggcacag aattga Gene symbol: LNX Entrez Gene ID: 84708 dna/ac: BCO22983.1 sequence: (SEQ ID NO: 14) atgaaccagccagagtctgccaacgatcctgaacccctgtgtgcagtg tgtggccaagcccactccttggaggaaaaccacttctacagctatcca gaggaagtggatgatgacctcatctgccacatctgcctgcaggattgc tggacccectggacactccgtgtggacacacctactgcaccctctgcc tcaccaacttcctggtggagaaggacttctgtcccatggaccgcaagc ctctggttctgcagcactgcaagaagtccagcatcctggtcaacaaac tcctcaacaagctactggtgacctgcccattcagggagcactgcaccc aggtgttgcagcgctgtgacctcgagcatcactttcaaaccagctgta aaggtgcctcccactacggcctgaccaaagataggaagaggcgctcac aagatggctgtccagacggctgtgcgagcctcacagccacggctccct ccccagaggtttctgcagctgccaccatctccttaatgacagacgagc ctggcctagacaaccctgcctacgtgtcctcggcagaggacgggcagc cagcaatcagcccagtggactctggccggagcaaccgaactagggcac ggccctttgagagatccactattagaagcagatcatttaaaaaaataa atcgagctttgagtgttcttcgaaggacaaagagcgggagtgcagttg ccaaccatgccgaccagggcagggaaaattctgaaaacaccactgccc ctgaagtctttccaaggttgtaccacctgattccagatggtgaaatta ccagcatcaagatcaatcgagtagatcccagtgaaagcctctctatta ggctggtgggaggtagcgaaaccccactggtccatatcattatccaac acatttatcgtgatggggtgatcgccagagacggccggctactgccag gagacatcattctaaaggtcaacgggatggacatcagcaatgtccctc acaactacgctgtgcgtctcctgcggcagccctgccaggtgctgtggc tgactgtgatgcgtgaacagaagttccgcagcaggaacaatggacagg ccccggatgcctacagaccccgagatgacagctttcatgtgattctca acaaaagtagccccgaggagcagcttggaataaaactggtgcgcaagg tggatgagcctggggttttcatcttcaatgtgctggatggcggtgtgg catatcgacatggtcagcttgaggagaatgaccgtgtgttagccatca atggacatgatcttcgatatggcagcccagaaagtgcggctcatctga ttcaggccagtgaaagacgtgttcacctcgtcgtgtcccgccaggttc ggcagcggagccctgacatctttcaggaagccggctggaacagcaatg gcagctggtccccagggccaggggagaggagcaacactcccaagcccc tccatcctacaattacttgtcatgagaaggtggtaaatatccaaaaag accccggtgaatctctcggcatgaccgtcgcagggggagcatcacata gagaatgggatttgcctatctatgtcatcagtgttgagcccggaggag tcataagcagagatggaagaataaaaacaggtgacattttgttgaatg tggatggggtcgaactgacagaggtcagccggagtgaggcagtggcat tattgaaaagaacatcatcctcgatagtactcaaagctttggaagtca aagagtatgagccccaggaagactgcagcagcccagcagccctggact ccaaccacaacatggccccacccagtgactggtccccatcctgggtca tgtggctggaattaccacggtgcttgtataactgtaaagatattgtat tacgaagaaacacagctggaagtctgggcttctgcattgtaggaggtt atgaagaatacaatggaaacaaaccttttttcatcaaatccattgttg aaggaacaccagcatacaatgatggaagaattagatgtggtgatattc ttcttgctgtcaatggtagaagtacatcaggaatgatacatgcttgct tggcaagactgctgaaagaacttaaaggaagaattactctaactattg tttcttggcctggcacttttttatag Gene symbol: MAP1S Entrez Gene ID: 55201 dna/ac: BC006358.1 sequence: (SEQ ID NO: 15) atgggcgtgggccggctggacatgtatgtgctgcacccgccctccgcc ggcgccgagcgcacgctggcctgtgtgtgcgccctgctggtgtggcac cccgccggccccggcgagaaggtggtgcgcgtgctgttccccggttgc accccgcccgcctgcctcctggacggcctggtccgcctgcagcacttg aggttcctgcgagagcccgtggtgacgccccaggacctggaggggccg gggcgagccgagagcaaagagagcgtgggctcccgggacagctcgaag agagagggcctcctggccacccaccctagacctggccaggagcgccct ggggtggcccgcaaggagccagcacgggctgaggccccacgcaagact gagaaagaagccaagaccccccgggagttgaagaaagaccccaaaccg agtgtctcccggacccagccgcgggaggtgcgccgggcagcctcttct gtgcccaacctcaagaagacgaatgcccaggcggcacccaagccccgc aaagcgcccagcacgtcccactctggcttcccgccggtggcaaatgga ccccgcagcccgcccagcctccgatgtggagaagccagcccccccagt gcggcctgcggctctccggcctcccagctggtggccacgcccagcctg gagctggggccgatcccagccggggaggagaaggcactggagctgcct ttggccgccagctcaatcccaaggccacgcacaccctcccctgagtcc caccggagccccgcagagggcagcgagcggctgtcgctgagcccactg cggggcggggaggccgggccagacgcctcacccacagtgaccacaccc acggtgaccacgccctcactacccgcagaggtgggctccccgcactcg accgaggtggacgagtccctgtcggtgtcctttgagcaggtgctgccg ccatccgcccccaccagtgaggctgggctgagcctcccgctgcgtggc ccccgggcgcggcgctcggcttccccacacgatgtggacctgtgcctg gtgtcaccctgtgaatttgagcatcgcaaggcggtgccaatggcaccg gcacctgcgtcccccggcagctcgaatgacagcagtgcccggtcacag gaacgggcaggtgggctgggggccgaggagacgccacccacatcggtc agcgagtccctgcccaccctgtctgactcggatcccgtgcccctggcc cccggtgcggcagactcagacgaagacacagagggctttggagtccct cgccacgaccctttgcctgaccccctcaaggtccccccaccactgcct gacccatccagcatctgcatggtggaccccgagatgctgccccccaag acagcacggcaaacggagaacgtcagccgcacccggaagcccctggcc cgccccaactcacgcgctgccgcccccaaagccactccagtggctgct gccaaaaccaaggggcttgctggtggggaccgtgccagccgaccactc agtgcccggagtgagcccagtgagaagggaggccgggcacccctgtcc agaaagtcctcaacccccaagactgccactcgaggcccgtcggggtca gccagcagccggcccggggtgtcagccaccccacccaagtccccggtc tacctggacctggcctacctgcccagcgggagcagcgcccacctggtg gatgaggagttcttccagcgcgtgcgcgcgctctgctacgtcatcagt ggccaggaccagcgcaaggaggaaggcatgcgggccgtcctggacgcg ctactggccagcaagcagcattgggaccgtgacctgcaggtgaccctg atccccactttcgactcggtggccatgcatacgtggtacgcagagacg cacgcccggcaccaggcgctgggcatcacggtgttgggcagcaacagc atggtgtccatgcaggatgacgccttcccggcctgcaaggtggagttc tag Gene symbol: NDUFV3 Entrez Gene ID: 4731 dna/ac: BCO21217.2 sequence: (SEQ ID NO: 16) atggctgccccgtgtttgctgcggcaaggacgagccggggcgctgaag actatgaccaggaagcccaggtgtttcgaggacttgcttctacggttt attgtctgcggaatcagggaagagtgaaaagggtcagccacagaattc caagaagcaaagtccaccaaaaaatgtagtggaaccaaaggagagggg caagacctagccacccagacagcagctgaattgtetaaaaacttatct tcacccagttcttacccgccagctgtgaataagggcaggaaggtagct agtcccagtcccagtggcagcgtgctattcacagatgaaggggttccg aaatttttgtcaagaaagactttggtagagtttccacagaaagttctg tctccattcagaaaacagggctctgattcagaagctcgtcaggtgggt cggaaagtgacgtcgccttcgtcttcatcctcgtccagctcctctgat tctgaatctgatgatgaggctgacgtttcagaggtcactcctcgagtg gtgagcaaaggcagaggggggcttcgaaaaccagaggcctctcattcc tttgaaaacagagccccccgagttacagtatcagcaaaagagaaaacc ttgctgcagaagccgcatgtggacattactgatccagagaagccccac cagccaaagaagaaagggtcccctgctaagccatcagaaggcagggaa aatgcgagaccaaaaaccacaatgcccagatctcaagtagatgaagag tttttgaagcaaagtttaaaggaaaaacaattgcagaaaacatttaga ttaaatgaaatagataaagaaagccaaaagccatttgaagttaaagga cccttacctgtccacacaaaatcagggttgtctgcgccaccgaagggc agcccagcgcctgctgtgttggcagaagaggccagagcagaggggcag ctgcaagccagtcctcctggggcggcagaggggcatctggaaaaaccc gtgccagagccccagcgcaaggcggcccctcccctgcccagaaaggaa acctcagggacgcagggaatagaaggccacctgaagggtggacaggca atcgtggaagatcagataccaccaagcaatttggagacagttcctgtt gagaataaccacggtttccatgaaaagacagcagcgctgaagcttgag gccgagggcgaggccatggaagatgcagccgcgccagggaacgaccga ggcggcacacaggagccagccccagtgcctgctgagccgtttgacaac actacctacaagaacctgcagcatcatgactacagcacgtacaccttc ttagacctcaacctcgaactctcaaaattcaggatgcctcagccctcc tcaggccgggagtcacctcgacactga Gene symbol: PABPC1 Entrez Gene ID: 26986 dna/ac: BC015958.2 sequence: (SEQ ID NO: 17) atgaaccccagtgcccccagctaccccatggcctcgctctacgtgggg gacctccaccccgacgtgaccgaggcgatgctctacgagaagttcagc ccggccgggcccatcctctccatccgggtctgcagggacatgatcacc cgccgctccttgggctacgcgtatgtgaacttccagcagccggcggac gcggagcgtgattggacaccatgaattttgatgttataaagggcaagc cagtacgcatcatgtggtctcagcgtgatccatcacttcgcaaaagtg gagtaggcaacatattcattaaaaatctggacaaatccattgataata aagcactgtatgatacattttctgatttggtaacatcctttcatgtaa ggtggtttgtgatgaaaatggttccaagggctatggatttgtacactt tgagacgcaggaagcagctgaaagagctattgaaaaaatgaatggaat gctcctaaatgatcgcaaagtatttgaggacgatttaagtctcgtaaa gaacgagaagctgaacttggagctagggcaaaagaattcaccaatgtt tacatcaagaattttggagaagacatggatgatgagcgccttaaggat ctctttggcaagtttgggcctgccttaagtgtgaaagtaatgactgat gaaagtggaaaatccaaaggatttggatttgtaagattgaaaggcatg aagatgcacagaaagctgtggatgagatgaacggaaaggagctcaatg gaaaacaaatttatgttggtcgagctcagaaaaaggtggaacggcaga cggaacttaagcgcaaatttgaacagatgaaacaagataggatcacca gataccagggtgttaatctttatgtgaaaaatcttgatgatggtattg atgatgaacgtctccggaaagagttttctccatttggtacaatcacta gtgcaaaggttatgatggagggtggtcgcagcaaagggtttggttttg tatgtttacctccccagaagaagccactaaagcagttacagaaatgaa cggtagaattgtggccacaaagccattgtatgtagctttagctcagcg caaagaagagcgccaggctcacctcactaaccagtatatgcagagaat ggcaagtgtacgagctgttcccaaccctgtaatcaacccctaccagcc agcacctccttcaggttacttcatggcagctatcccacagactcagaa ccgtgctgcatactatcctcctagccaaattgctcaactaagaccaag tcctcgctggactgctcagggtgccagacctcatccattccaaaatat gcccggtgctatccgcccagctgctcctagaccaccatttagtactat gagaccagcttcttcacaggttccacgagtcatgtcaacacagcgtgt tgctaacacatcaacacagacaatgggtccacgtectgcagctgcagc cgctgcagctactcctgctgtccgcaccgttccacagtataaatatgc tgcaggagttcgcaatcctcagcaacatcttaatgcacagccacaagt tacaatgcaacagcctgctgttcatgtacaaggtcaggaacctttgac tgcttccatgttggcatctgcccctcctcaagagcaaaagcaaatgtt gggtgaacggctgtttcctcttattcaagccatgcaccctactcttgc tggtaaaatcactggcatgttgttggagattgataattcagaacttct tcatatgctcgagtctccagagtcactccgttctaaggttgatgaagc tgtagctgtactacaagcccaccaagctaaagaggctgcccagaaagc agttaacagtgccaccggtgttccaactgtttaa Gene symbol: PHLDA1 Entrez Gene ID: 22822 dna/ac: BC018929.2 sequence: (SEQ ID NO: 18) atgctggagagtagcggctgcaaagcgctgaaggagggcgtgctggag aagcgcagcgacgggttgttgcagctctggaagaaaaagtgttgcatc ctcaccgaggaagggctgctgcttatcccgcccaagcagctgcaacac cagcagcagcagcaacagcagcagcagcagcagcaacaacagcccggg caggggccggccgagccgtcccaacccagtggccccgctgtcgccagc ctcgagccgccggtcaagctcaaggaactgcacttctccaacatgaag accgtggactgtgtggagcgcaagggcaagtacatgtacttcactgtg gtgatggcagagggcaaggagatcgactttcggtgcccgcaagaccag ggctggaacgccgagatcacgctgcagatggtgcagtacaagaatcgt caggccatcctggcggtcaaatccacgcggcagaagcagcagcacctg gtccagcagcagcccccctcgcagccgcagccgcagccgcagctccag ccccaaccccagcctcagcctcagccgcaaccccagccccaatcacaa ccccagcctcagccccaacccaagcctcagccccagcagctccacccg tatccgcatccacatccacatccacactctcatcctcactcgcaccca caccctcacccgcacccgcatccgcaccaaataccgcacccacaccca cagccgcactcgcagccgcacgggcaccggcttctccgcagcacctcc aactctgcctga Gene symbol: PIAS2 Entrez Gene ID: 9063 dna/ac: BC015190.1 sequence: (SEQ ID NO: 19) atggcggatttcgaagagttgaggaatatggtttctagttttagggtt tctgaactacaagtattactaggattgctggacggaataaaagtggac gcaagcatgacctcctgatgagggcgctgcatttattgaagagcggct gcagccctgcggttcagattaaaatccgagaattgtatagacgccgat atccacgaactcttgaaggactttctgatttatccacaatcaaatcat cggattcagtttggatggtggctcatcacctgtagaacctgacttggc cgtggctggaatccactcgttgccttccacttcagttacacctcactc accatcctctcctgttggttctgtgctgcttcaagatactaagcccac atttgagatgcagcagccatctcccccaattcctcctgtccatcctga tgtgcagttaaaaaatctgccatttatgatgtccttgatgactcatca agcccacgagtttagttcaaagcagtattcagcgatttcaagagaagt tttttatttttgctttgacacctcaacaagttagagagatatgcatat ccagggattttttgccaggtggtaggagagattatacagtccaagttc agttgagactttgcctggcagagacaagttgccctcaagaagataact atccaaatagtctatgtataaaagtaaatgggaagctatttcctttgc ctggctatgcaccaccgcctaaaaatgggattgaacagaagcgccctg gacgccccttgaatattacatctttagttaggttatcttcagctgtgc caaaccaaatttccatttcttgggcatcagaaattgggaagaattact ctatgtctgtatatcttgtacggcagcttacatcagccatgttattac agagattaaaaatgaaaggtattagaaaccctgatcattccagagcac taattaaagaaaaacttactgcagatcctgatagtgaaattgctacaa ctagccttcgggtatccttgatgtgccctttaggaaaaatgaggctga caatcccatgccgtgcagtgacttgtacacatctgcagtgttttgatg ctgccctctatctacaaatgaatgagaaaaagcccacctggatttgtc ctgtgtgtgacaaaaaagctgcctatgaaagtctaatattagatgggc tttttatggaaattctcaatgactgttctgatgtagatgagatcaaat tccaagaagatggttcttggtgtccaatgagaccgaagaaagaagcta tgaaagtatccagccaaccgtgtacaaaaatagaaagttcaagcgtcc tcagtaagccttgttcagtgactgtagccagtgaggcaagcaagaaga aagtagatgttattgatcttacaatagaaagctcttctgacgaagagg aagaccctcctgccaaaaggaaatgcatctttatgtcagaaacacaaa gcagcccaaccaaaggggttctcatgtatcagccatcttctgtaaggg tgcccagtgtgacttcggttgatcctgctgctattccgccttcattaa cagactactcagtaccattccaccatacgccaatatcaagcatgtcat cagatttgccaggagaacaaagaagaaatgatattaataatgaaccga agcttggaacatcttctgatactgtgcaacagtga Gene symbol: PRKCBP1 Entrez Gene ID: 23613 dna/ac: BC030721.1 sequence: (SEQ ID NO: 20) atgacaatgctcaccattgaacagttatcctacctgctcaagtttgcc attcagaaaatgaaacagccagggacagatgcattccagaagcccgtt ccattggaacagcaccctgactatgcggaatacatcttccatccaatg gacctttgtacattggaaaagaatgcgaaaaagaaaatgtatggctgc acagaagccttcctggctgatgcaaagtggattttgcacaactgcatc atttataatgggggaaatcacaaattgacgcaaatagcgaaagtagtc atcaaaatctgtgaacatgagatgaatgaaatcgaagtatgtccagaa tgttatctagctgcttgccaaaaacgagataactggttttgtgagcct tgtagcaatccacatcctttggtctgggccaaactgaaggggtttcca ttctggcctgcaaaagctctaagggataaagacgggcaggtcgatgcc cgattctttggacaacatgacagggcctgggttccaataaataattgc tacctcatgtctaaagaaattcctttttctgtgaaaaagactaagagc atcttcaacagtgccatgcaagagatggaggtttacgtggagaacatc cgcaggaagtttggggtttttaattactctccatttaggacaccctac acacccaacagccagtatcaaatgctgctcgatcccaccaaccccagc gccggcactgccaagatagacaagcaggagaaggtcaagctcaacttt gacatgacggcatcccccaagatcctgatgagcaagcctgtgctgagt gggggcacaggccgccggatttccttgtcggatatgccgcgctccccc atgagcacaaactcttctgtgcacacgggctccgacgtggagcaggat gctgagaagaaggccacgtcgagccacttcagtgcgagcgaggagtcc atggacttcctggataagagcacagcttcaccagcctccaccaagacg ggacaagcagggagtttatccggcagcccaaagcccttctctcctcaa ctgtcagctcctatcacgacgaaaacggacaaaacctccaccaccggc agcatcctgaatcttaacctggatcgaagcaaagctgagatggatttg aaggagctgagcgagtcggtccagcaacagtccacccctgttcctctc atctctcccaagcgccagattcgtagcaggttccagctgaatcttgac aagaccatagagagttgcaaagcacaattaggcataaatgaaatctcg gaagatgtctatacggccgtagagcacagcgattcggaggattctgag aagtcagatagtagcgatagtgagtatatcagtgatgatgagcagaag tctaagaacgagccagaagacacagaggacaaagaaggttgtcagatg gacaaagagccatctgctgttaaaaaaaagcccaagcctacaaaccca gtggagattaaagaggagctgaaaagcacgtcaccagccagcgagaag gcagaccctggagcagtcaaggacaaggccagccctgagcctgagaag gacttttccgaaaaggcaaaaccttcacctcaccccataaaggataaa ctgaagggaaaagatgagacggattccccaacagtccatttgggcctg gactctgattcagagagcgaacttgtcatagatttaggagaagaccat tctgggcgggagggtcgaaaaaataagaaggaacccaaagaaccatct cccaaacaggatgttgtaggtaaaactccaccatccacgacggtgggc agccattctcccccggaaacaccggtgctcacccgctcttccgcccaa acttccgcggctggcgccacagccaccaccagcacgtcctccacggtc accgtcacggccccggcccccgccgccacaggaagcccagtgaaaaag cagaggccgcttttaccgaaggagactgccccggccgtgcagcgggtc gtgtggaactcatcaactgtccagcagaaggagatcacacagagccca tccacgtccaccatcaccctggtgaccagcacacagtcatcgcccctg gtcaccagctcggggtccatgagcacccttgtgtcctcagtcaacgct gacctgcccatcgccactgcctcagctgatgtcgccgctgatattgcc aagtacactagcaaaatgatggatgcaataaaaggaacaatgacagaa atatacaacgatctttctaaaaacactactggaagcacaatagctgag attcgcaggctgaggatcgagatagagaagctccagtggctgcaccag caagagctctccgaaatgaaacacaacttagagctgaccatggcggag atgcggcagagcctggagcaggagcgggaccggctcatcgccgaggtg aagaagcagctggagttggagaagcagcaggcggtggatgagaccaag aagaagcagtggtgcgccaactgcaagaaggaggccatcttttactgc tgttggaacaccagctactgtgactacccctgccagcaagcccactgg cctgagcacatgaagtcctgcacccagtcagctactgctcctcagcag gaagcggatgctgaggtgaacacagaaacactaaataagtcctcccag gggagctcctcgagcacacaatcagcaccttcagaaacggccagcgcc tccaaagagaaggagacgtcagctgagaaaagcaaggagagtggctcg acccttgacctttctggctccagagagacgccctcctccattctctta ggctccaaccaaggctctgttagcaaaaggtgtgacaagcaacctgcc tatgccccaaccaccacagaccaccagccgcaccccaactaccccgcc cagaagtaccattcccggagtaataaatccagttggagcagcagtgat gagaagaggggatcgacacgttccgatcacaacaccagtaccagcacg aagagcctcctcccgaaagagtctcggctggacaccttctgggactag Gene symbol: PRKRA Entrez Gene ID: 8575 dna/ac: BC009470.1 sequence: (SEQ ID NO: 21) atgtcccagagcaggcaccgcgccgaggccccgccgctggagcgcgag gacagtgggaccttcagtttggggaagatgataacagctaagccaggg aaaacaccgattcaggtattacacgaatacggcatgaagaccaagaac atcccagtttatgaatgtgaaagatctgatgtgcaaatacacgtgccc actttcaccttcagagtaaccgttggtgacataacctgcacaggtgaa ggtacaagtaagaagctggcgaaacatagagctgcagaggctgccata aacattttgaaagccaatgcaagtatttgctttgcagttcctgacccc ttaatgcctgacccttccaagcaaccaaagaaccagcttaatcctatt ggttcattacaggaattggctattcatcatggctggagacttcctgaa tataccctttcccaggagggaggacctgctcataagagagaatatact acaatttgcaggctagagtcatttatggaaactggaaagggggcatca aaaaagcaagccaaaaggaatgctgctgagaaatttcttgccaaattt agtaatatttctccagagaaccacatttctttaacaaatgtagtagga cattctttaggatgtacttggcattccttgaggaattctcctggtgaa aagatcaacttactgaaaagaagcctccttagtattccaaatacagat tacatccagctgcttagtgaaattgccaaggaacaagttttaatataa catatttggatatagatgaactgagcgccaatggacaatatcaatgtc ttgctgaactgtccaccagccccatcacagtctgtcatggctccggta tctcctgtggcaatgcacaaagtgatgcagctcacaatgctttgcagt atttaaagataatagcagaaagaaagtaa Gene symbol: PSME3 Entrez Gene ID: 10197 dna/ac: BC001423.2 sequence: (SEQ ID NO: 22) atggcctcgttgctgaaggtggatcaggaagtgaagctcaaggttgat tctttcagggagcggatcacaagtgaggcagaagacttggtggcaaat tttttcccaaagaagttattagaacttgatagttttctgaaggaacca atcttaaacatccatgacctaactcagatccactctgacatgaatctc ccagtccctgaccccattcttctcaccaatagccatgatggactggat ggtcccacttataagaagcgaaggttggatgagtgtgaagaagccttc caaggaaccaaggtgtttgtgatgcccaatgggatgctgaaaagcaac cagcagctggtggacattattgagaaagtgaaacctgagatccggctg ttgattgagaaatgtaacacggtcaaaatgtgggtacagctcctgatt cccaggatagaagatggaaacaactttggggtgtccattcaggaggaa acagttgcagagctaagaactgttgagagtgaagctgcatcttatctg gaccagatttctagatattatattacaagagccaaattggtttctaaa atagctaaatatccccatgtggaggactatcgccgcaccgtgacagag attgatgagaaagaatatatcagccttcggctcatcatatcagagctg aggaatcaatatgtcactctacatgacatgatcctgaaaaatatcgag aagatcaaacggccccggagcagcaatgcagagactctgtactga Gene symbol: RAB11FIP3 Entrez Gene ID: 9727 dna/ac: BC051360.1 sequence: (SEQ ID NO: 23) atggcgtcggccccgccggcctcgcccccgggctcggagccgccgggg cccgacccggagccgggcgggccggacgggccgggggcggcgcaactg gctccgggccctgcggagctacgcctcggagcgcccgtcggcggcccc gacccgcagtccccgggcctggatgagcctgcgcccggggccgctgca gatggcggggcgcgttggagcgccgggccggccccggggctggaggga ggcccgcgagaccccgggccgtccgccccgccgccgcgctccggcccg cgggggcagcttgcgagccccgacgccccgggcccagggccgcgctcc gaagcgccgcttccagaactcgacccgttgttctcctggactgaggag cccgaggagtgtggccccgcgagctgcccggagagcgcgcctttccgc ttgcaggggtccagcagcagccaccgagcgcggggcgaggtcgacgtc ttctctcccttccccgcgcccacggcgggcgagctggcgctggagcaa ggtcccgggtccccgccgcagccctcggacctcagccagacccacccc cttccgagcgagcccgtggggagtcaggaggacggcccccgcctccga gccgtgttcgatgccctggacggggatggggacggtttcgtccgcatc gaggacttcatccagtttgctacggtctacggggcagagcaggtgaag gacttaactaagtacttggatcccagtgggctcggcgtgatcagcttt gaagacttctaccaagggatcacagccatcagaaacggagatcctgat ggccagtgctacggtggtgtcgcttctgcccaagatgaggagcccctg gcctgcccggacgagttcgatgacttcgtcacctatgaggccaacgag gtgacggacagcgcgtacatgggctccgagagcacctacagtgagtgt gagaccttcacggacgaggacaccagcaccctggtgcaccctgagctg caacctgaaggggacgcagacagtgccggcggctcggccgtgccctct gagtgcctggacgccatggaggagcccgaccatggtgccctgctgctg ctcccaggcaggcctcacccccatggccagtctgtcatcacggtgatc gggggcgaggagcactttgaggactacggtgaaggcagtgaggcggag ctgtccccagagaccctatgcaacgggcagctgggctgcagtgacccc gctttcctcacgcccagtccgacaaagcggctctccagcaagaaggtg gcaaggtacctgcaccagtcaggggccctgaccatggaggccctggag gacccttcccccgagctcatggagggcccagaggaggacattgctgac aaggttgtcttcctggaaaggcgtgtgctggagctggaaaaggacacg gcagccaccggtgagcaacacagccgcctgaggcaggagaacctgcag ctggtgcacagagcaaacgccctggaggagcagctgaaggagcaggag ctgagagcctgcgagatggtcctggaagagacccggcgtcagaaggag ctcctgtgcaagatggagagggagaagagcattgagatcgagaacctg cagaccaggctacagcaactggacgaggagaacagtgaactccggtcc tgcacgccctgtctgaaggccaacattgagcgtctggaggaggagaag cagaagctgttggatgagatagagtcgctgacgctgcggctcagtgaa gagcaggagaacaagaggagaatgggggacaggctgagtcacgagagg caccagttccagagggacaaggaggccacccaggagctgatcgaggac ctccgaaagcagctggagcacctgcagctcctcaagctggaggccgag cagcggcggggccgcagcagcagcatgggcctgcaggagtaccacagc cgcgcccgggagagcgagctggagcaggaggtccgcaggctgaagcag gacaaccgcaacctgaaggagcagaacgaggagctgaacgggcagatc attaccctcagcatccagggcgccaagagcctcttctccacagccttc tctgagtccctggctgcagagatcagctccgtctcccgagatgagctc atggaggcgattcagaagcaggaggagatcaacttccgcctgcaggac tacatcgacaggatcatcgtggccatcatggagaccaacccgtccatc ctggaggtcaagtag Gene symbol: RALBP1 Entrez Gene ID: 10928 dna/ac: BC013126.1 sequence: (SEQ ID NO: 24) atgactgagtgcttcctgccccccaccagcagccccagtgaacaccgc agggtggagcatggcagcgggcttacccggacccccagctctgaagag atcagccctactaagtttcctggattgtaccgcactggcgagccctca cctccccatgacatcctccatgagcctcctgatgtagtgtctgatgat gagaaagatcatgggaagaaaaaagggaaatttaagaaaaaggaaaag aggactgaaggctatgcagcctttcaggaagatagctctggagatgag gcagaaagtccttctaaaatgaagaggtccaagggaatccatgttttc aagaagcccagcttttctaaaaagaaggaaaaggattttaaaataaaa gagaaacccaaagaagaaaagcataaagaagaaaagcacaaagaagaa aaacataaagagaagaagtcaaaagacttgacagcagctgatgttgtt aaacagtggaaggaaaagaagaaaaagaaaaagccaattcaggagcca gaggtgcctcagattgatgttccaaatctcaaacccatttttggaatt cattggctgatgcagtagagaggaccatgatgtatgatggcattcggc tgccagccgttttccgtgaatgtatagattacgtagagaagtatggca tgaagtgtgaaggcatctacagagtatcaggaattaaatcaaaggtgg atgagctaaaagcagcctatgaccgggaggagtctacaaacttggaag actatgagcctaacactgtagccagtttgctgaagcagtatttgcgag accttccagagaatttgataccaaagagcttatgcccagatttgaaga ggcttgtgggaggaccacggagactgagaaagtgcaggaattccagcg tttactcaaagaactgccagaatgtaactatcttctgatttatggctc attgtgcacatggaccatgtcattgcaaaggaactggaaacaaaaatg aatatacagaacatttctatagtgctcagcccaactgtgcagatcagc aatcgagtcctgtatgtgtttttcacacatgtgcaagaactattggaa atgtggtactaaagcaagtgatgaaacctctgcgatggtctaacatgg ccacgatgcccacgctgccagagacccaggcgggcatcaaggaggaga tcaggagacaggagtttatttgaattgtttacatcgagatctgcaggg tgggataaaggatttgtctaaagaagaaagattatgggaagtacaaag aattttgacagccctcaaaagaaaactgagagaagctaaaagacagga gtgtgaaaccaagattgcacaagagatagccagtattcaaaagaggat gatccaaagaagagatgaatgaaaatgaagaagttataaatattctcc ttgctcaggagaatgagatcctgactgaacaggaggagctcctggcca tggagcagtttctgcgccggcagattgcctcagaaaaagaagagattg aacgcctcagagctgagattgctgaaattcagagtcgccagcagcacg gccgaagtgagactgaggagtactcctccgagagcgagagcgagagtg aggatgaggaggagctgcagatcattctggaagacttacagagacaga acgaagagctggaaataaagaacaatcatttgaatcaagcaattcatg aggagcgcgaggccatcatcgagctgcgcgtgcagctgcggctgctcc agatgcagcgagccaaggccgagcagcaggcgcaggaggacgaggagc ctgagtggcgcgggggtgccgtccagccgcccagagacggcgtccttg agccaaaagcagctaaagagcagccaaaggcaggcaaggagccggcaa agccatcgcccagcagggataggaaggagacgtccatctga Gene symbol: RAN Entrez Gene ID: 5901 dna/ac: BC014518.2 sequence: (SEQ ID NO: 25) atggctgcgcagggagagccccaggtccagttcaaacttgtattggtt ggtgatggtggtactggaaaaacgaccttcgtgaaacgtcatttgact ggtgaatttgagaagaagtatgtagccaccttgggtgttgaggttcat cccctagtgttccacaccaacagaggacctattaagttcaatgtatgg gacacagccggccaggagaaattcggtggactgagagatggctattat atccaagcccagtgtgccatcataatgtttgatgtaacatcgagagtt acttacaagaatgtgcctaactggcatagagatctggtacgagtgtgt gaaaacatccccattgtgttgtgtggcaacaaagtggatattaaggac aggaaagtgaaggcgaaatccattgtcttccaccgaaagaagaatctt cagtactacgacatttctgccaaaagtaactacaactttgaaaagcca tcctctggcttgctaggaagctcattggagaccctaacttggaatttg ttgccatgcctgctctcgccccaccagaagttgtcatggacccagctt tggcagcacagtatgagcacgacttagaggttgctcagacaactgctc tcccggatgaggatgatgacctgtga Gene symbol: RARA Entrez Gene ID: 5914 dna/ac: BC008727.2 sequence: (SEQ ID NO: 26) atggccagcaacagcagctcctgcccgacacctgggggcgggcacctc aatgggtacccggtgcctccctacgccttcttcttcccccctatgctg ggtggactctccccgccaggcgctctgaccactctccagcaccagctt ccagttagtggatatagcacaccatccccagccaccattgagacccag agcagcagttctgaagagatagtgcccagccctccctcgccaccccct ctaccccgcatctacaagccttgctttgtctgtcaggacaagtcctca ggctaccactatggggtcagcgcctgtgagggctgcaagggcttcttc cgccgcagcatccagaagaacatggtgtacacgtgtcaccgggacaag aactgcatcatcaacaaggtgacccggaaccgctgccagtactgccga ctgcagaagtgctttgaagtgggcatgtccaaggagtctgtgagaaac gaccgaaacaagaagaagaaggaggtgcccaagcccgagtgctctgag agctacacgctgacgccggaggtgggggagctcattgagaaggtgcgc aaagcgcaccaggaaaccttccctgccctctgccagctgggcaaatac actacgaacaacagctcagaacaacgtgtctctctggacattgacctc tgggacaagttcagtgaactctccaccaagtgcatcattaagactgtg gagttcgccaagcagctgcccggcttcaccaccctcaccatcgccgac cagatcaccctcctcaaggctgcctgcctggacatcctgatcctgcgg atctgcacgcggtacacgcccgagcaggacaccatgaccttctcggac gggctgaccctgaaccggacccagatgcacaacgctggcttcggcccc ctcaccgacctggtctttgccttcgccaaccagctgctgcccctggag atggatgatgcggagacggggctgctcagcgccatctgcctcatctgc ggagaccgccaggacctggagcagccggaccgggtggacatgctgcag gagccgctgctggaggcgctaaaggtctacgtgcggaagcggaggccc agccgcccccacatgttccccaagatgctaatgaagattactgacctg cgaagcatcagcgccaagggggctgagcgggtgatcacgctgaagatg gagatcccgggctccatgccgcctctcatccaggaaatgttggagaac tcagagggcctggacactctgagcggacagccggggggtggggggcgg gacgggggtggcctggcccccccgccaggcagctgtagccccagcctc agccccagctccaacagaagcagcccggccacccactccccgtga Gene symbol: RBMS1 Entrez Gene ID: 5937 dna/ac: BC012992.2 sequence: (SEQ ID NO: 27) atgggcaaagtgtggaaacagcagatgtaccctcagtacgccacctac tattacccccagtatctgcaagccaagcagtctctggtcccagcccac cccatggcccctcccagtcccagcaccaccagcagtaataacaacagt agcagcagtagcaactcaggatgggatcagctcagcaaaacgaacctc tatatccgaggactgcctccccacaccaccgaccaggacctggtgaag ctctgtcaaccatatgggaaaatagtctccacaaaggcaattttggat aagacaacaaacaaatgcaaaggttatggttttgtcgactttgacagc cctgcagcagctcaaaaagctgtgtctgccctgaaggccagtggggtt caagctcaaatggcaaagcaacaggaacaagatcctaccaacctctac atttctaatttgccactctccatggatgagcaagaactagaaaatatg ctcaaaccatttggacaagttatttctacaaggatactacgtgattcc agtggtacaagtcgtggtgttggctttgctaggatggaatcaacagaa aaatgtgaagctgttattggtcattttaatggaaaatttattaagaca ccaccaggagtttctgcccccacagaacctttattgtgtaagtttgct gatggaggacagaaaaagagacagaacccaaacaaatacatccctaat ggaagaccatggcatagagaaggagaggctggaatgacacttacttac gacccaactacagctgctatacagaacggattttatccttcaccatac agtattgctacaaaccgaatgatcactcaaacttctattacaccctat attgcatctcctgtatctgcctaccaggtgcaaagtccttcgtggatg caacctcaaccatatattctacagcaccctggtgccgtgttaactccc tcaatggagcacaccatgtcactacagcccgcatcaatgatcagccct ctggcccagcagatgagtcatctgtcactaggcagcaccggaacatac atgcctgcaacgtcagctatgcaaggagcctacttgccacagtatgca catatgcagacgacagcggttcclgttgaggaggcaagtggtcaacag caggtggctgtcgagacgtctaatgaccattctccatatacctttcaa cctaataagtaa Gene symbol: RDBP Entrez Gene ID: 7936 dna/ac: BC050617.2 sequence: (SEQ ID NO: 28) atgttggtgataccccccggactgagcgaggaagaggaggctctgcag aagaaattcaacaagctcaagaaaaagaaaaaggcattgctggctctg aagaagcaaagtagcagcagcacaaccagccaaggtggtgtcaaacgc tcactatcagagcagcctgtcatggacacagccacagcaacagagcag gcaaagcagctggtgaagtcaggagccatcagtgccatcaaggctgag accaagaactcaggcttcaagcgttctcgaacccttgaggggaagtta aaggaccccgagaagggaccagtccccactttccagccgttccagagg agcatatctgctgatgatgacctgcaagagtcatccagacgtccccag aggaaatctctgtatgagagctttgtgtcttctagtgatcgacttcga gaactaggaccagatggagaagaggcagagggcccaggggctggtgat ggtccccctcgaagctttgactggggctatgaagaacgcagtggtgcc cactcctcagcctcccctccccgaagccgcagccgggaccgcagccat gagaggaaccgggacagagaccgagatcgggagcgggatcgagaccgg gatcgagacagagacagagagcgggacagggatcgggatcgggatcga gatcgagaccgggaacgggacagggatcgggagcgggatcgagaccga gaccgagagggtcctttccgcaggtcggattcattccctgaacggcga gcccctaggaaagggaatactctctatgtatatggagaagacatgaca cccacccttctccgtggggccttctctccttttggaaacatcattgac ctctccatggacccacccagaaactgtgccttcgtcacctatgaaaag atggagtcagcagatcaggccgttgctgagctcaacgggacccaggtg gagtctgtacagctcaaagtcaacatagcccgaaaacagcccatgctg gatgccgctactggcaagtctgtctggggctccctcgctgtccagaac agccctaagggttgccaccgggacaagaggacccagattgtctacagt gatgacgtctacaaggaaaaccttgtggatggcttctag Gene symbol: RNF12 Entrez Gene ID: 51132 dna/ac: BC013357.2 sequence: (SEQ ID NO: 29) atggaaaactcagattccaatgacaaaggaagtggtgatcagtctgca gcacagcgcagaagtcagatggaccgattggatcgagaagaagctttc tatcaatttgtaaataacctgagtgaagaagattataggcttatgaga gataacaatttgctaggcaccccaggtgaaagtactgaggaagagttg ctgagacgactacagcaaattaaagaaggcccaccaccgcaaaactca gatgaaaatagaggaggagactcttcagatgatgtgtctaatggtgac tctataatagactggcttaactctgtcagacaaactggaaatacaaca agaagtgggcaaagaggaaaccaatcttggagagcagtgagtcggact aatccaaacagtggtgatttcagattcagtttagagataaatgttaac cgtaataatgggagccaaaattcagagaatgaaaatgagccatctgca agacgttctagtggagaaaatgtggaaaacaacagccaaaggcaagtg gaaaacccacgatctgaatcaacatctgcaaggccatctagatcagaa cgaaattcaactgaagcgttaacagaggtcccacctaccagaggtcag aggagggcaagaagcaggagcccagaccatcggagaaccagagcaaga gctgaaagaagtaggtcacctctgcatccaatgagtgaaattccacga agatctcatcatagtatctcatctcagacttttgaacatcctttggta aatgagacggagggaagttctagaacccggcaccatgtgacattgagg cagcaaatatctgggcctgagttgctaagtagaggtctttttgcagct tctggaacaagaaatgcttctcaaggagcaggttcttcagacacagct gccagtggtgaatctacaggatcaggacagagacctccaaccatagtc cttgatcttcaagtaagaagagttcgtcctggagaatatcggcagaga gatagcatagccagcagaactcggtctaggtctcagacaccaaacaac actgtcacctatgaaagtgaacgaggaggttttaggcgtacattttca cgttctgagcgggcaggtgtgagaacctatgtcagtaccatcagaatt cccattcgtagaatcttaaatactggtttaagtgagactacatctgtt gcaattcagaccatgttaaggcagataatgacaggttttggtgagtta agctattttatgtacagtgatagcgactcagagcctactggctcagtc tcaaatcgaaatatggaaagggcagagtcacggagtggaagaggaggt tctggtggtggtagtagttctggttccagttcgagttccagttccagt tcgagttccagttccagttcaagttccagttccagtcctagttccagt tccggtggtgaaagttcagaaactagctcagatttatttgaaggcagt aatgaaggaagctcatcatcaggctcatcaggtgccaggcgagagggt cgacatagggccccagtcacatttgatgaaagtggctctttgcccttc cttagcctggctcagtttttcctcttaaatgaggatgatgatgaccaa cctagaggactcaccaaagaacagattgacaacttggcaatgagaagt tttggtgaaaatgatgcattaaaaacctgtagtgtttgcattacagaa tatacagaaggcaacaaacttcgtaaactaccttgttcccatgagtac catgtccactgcatcgatcgctggttatctgagaattctacctgtcct atttgtcgcagagcagtcttagcttctggtaacagagaaagtgttgtg taa Gene symbol: RPL30 Entrez Gene ID: 6156 dna/ac: BC032700.2 sequence: (SEQ ID NO: 30) atggtggccgcaaagaagacgaaaaagtcgctggagtcgatcaactct aggctccaactcgttatgaaaagtgggaagtacgtcctggggtacaag cagactctgaagatgatcagacaaggcaaagcgaaattggtcattctc gctaacaactgcccagctttgaggaaatctgaaatagagtactatgct atgttggctaaaactggtgtccatcactacagtggcaataatattggg cacagcatgcggaaaatactacagagtgtgcacactggctatcattga tccaggtgactctgacatcattagaagcatgccagaacagactggtga aaagtaa Gene symbol: RPL31 Entrez Gene ID: 6160 dna/ac: BC017343.2 sequence: (SEQ ID NO: 31) atggctcccgcaaagaagggtggcgagaagaaaaagggccgttctgcc atcaacgaagtggtaacccgagaatacaccatcaacattcacaagcgc atccatggagtgggcttcaagaagcgtgcacctcgggcactcaaagag attcggaaatttgccatgaaggagatgggaactccagatgtgcgcatt gacaccaggctcaacaaagctgtctgggccaaaggaataaggaatgtg ccataccgaatccgtgtgcggctgtccagaaaacgtaatgaggatgaa gattcaccaaataagctatatactttggttacctatgtacctgttacc actttcaaaaatctacagacagtcaatgtggatgagaactaa Gene symbol: RUFY1 Entrez Gene ID: 80230 dna/ac: BC032571.1 sequence: (SEQ ID NO: 32) atgatggaggagcgtgccaacctgatgcacatgatgaaactcagcatc aaggtgttgctccagtcggctctgagcctgggccgcagcctggatgcg gaccatgcccccttgcagcagttctttgtagtgatggagcactgcctc aaacatgggctgaaagttaagaagagttttattggccaaaataaatca ttctttggtcctttggagctggtggagaaactttgtccagaagcatca gatatagcgactagtgtcagaaatcttccagaattaaagacagctgtg ggaagaggccgagcgtggctttatcttgcactcatgcaaaagaaactg gcagattatctgaaagtgcttatagacaataaacatctcttaagcgag ttctatgagcctgaggctttaatgatggaggaagaagggatggtgatt gttggtctgctggtgggactcaatgttctcgatgccaatctctgcttg aaaggagaagacttggattctcaggttggagtaatagatttttccctc taccttaaggatgtgcaggatcttgatggtggcaaggagcatgaaaga attactgatgtccttgatcaaaaaaattatgtggaagaacttaaccgg cacttgagctgcacagttggggatcttcaaaccaagatagatggcttg gaaaagactaactcaaagcttcaagaagagctttcagctgcaacagac cgaatttgctcacttcaagaagaacagcagcagttaagagaacaaaat gaattaattcgagaaagaagtgaaaagagtgtagagataacaaaacag gataccaaagttgagctggagacttacaagcaaactcggcaaggtctg gatgaaatgtacagtgatgtgtggaagcagctaaaagaggagaagaaa gtccggttggaactggaaaaagaactggagttacaaattggaatgaaa accgaaatggaaattgcaatgaagttactggaaaaggacacccacgag aagcaggacacactagttgccctccgccagcagctggaagaagtcaaa gcgattaatttacagatgtttcacaaagctcagaatgcagagagcagt ttgcagcagaagaatgaagccatcacatcctttgaaggaaaaaccaac caagttatgtccagcatgaaacaaatggaagaaaggttgcagcactcg gagcgggcgaggcagggagctgaggagcggagccacaagctgcagcag gagctgggcgggaggatcggcgccctgcagctgcagctctcccagctg cacgagcaatgctcaagcctggagaaagaattgaaatcagaaaaagag caaagacaggctcttcagcgcgaattacagcacgagaaagacacttcc tctctactcaggatggagctgcaacaagtggaaggactgaaaaaggag ttgcgggagcncaggacgagaaggcagagctgcagaagatctgcgagg agcaggaacaagccctccaggaaatgggcctgcacctcagccagtcca agctgaagatggaagatataaaagaagtgaaccaggcactgaagggcc acgcctggctgaaagatgacgaagcgacacactgtaggcagtgtgaga aggagttctccatttcccggagaaagcaccactgccggaactgtggcc acatcttctgcaacacctgctccagcaacgagctggccctgccctcct accccaagccggtgcgagtgtgcgacagctgccacaccctgctcctgc agcgctgctcctccacggcctcctga Gene symbol: SMN1 Entrez Gene ID: 6606 dna/ac: BC000908.2 sequence: (SEQ ID NO: 33) atggcgatgagcagcggcggcagtggtggcggcgtcccggagcaggag gattccgtgctgttccggcgcggcacaggccagagcgatgattctgac atttgggatgatacagcactgataaaagcatatgataaagctgtggct tcatttaagcatgctctaaagaatggtgacatttgtgaaacttcgggt aaaccaaaaaccacacctaaaagaaaacctgctaagaagaataaaagc caaaagaagaatactgcagcttccttacaacagtggaaagttggggac aaatgttctgccatttggtcagaagacggttgcatttacccagctacc attgtttcaattgattttaagagagaaacctgtgttgtggtttacact ggatatggaaatagagaggagcaaaatctgtccgatctactttcccca atctgtgaagtagctaataatatagaacagaatgctcaagagaatgaa aatgaaagccaagtttcaacagatgaaagtgagaactccaggtctcct ggaaataaatcagataacatcaagcccaaatctgctccatggaactct tttctccctccaccaccccccatgccagggccaagactgggaccagga aagccaggtctaaaattcaatggcccaccaccgccaccgccaccacca ccaccccacttactatcatgctggctgcctccatttccttctggacca ccaataattcccccaccacctcccatatgtccagattctcttgatgat gctgatgctttgggaagtatgttaatttcatggtacatgagtggctat catactggctattatatggaaatgctggcatag Gene symbol: SNK Entrez Gene ID: 10769 dna/ac: BC013879.2 sequence: (SEQ ID NO: 34) atggagcttttgcggactatcacctaccagccagccgccagcaccaaa atgtgcgagcaggcgctgggcaagggttgcggagcagactcgaagaag aagcggccgccgcagccccccgaggaatcgcagccacctcagtcccag gcgcaagtgcccccggcggcccctcaccaccatcaccaccattcgcac tcggggccggagatctcgcggattatcgtcgaccccacgactgggaag cgctactgccggggcaaagtgctgggaaagggtggattgcaaaatgtt acgagatgacagatttgacaaataacaaagtctacgccgcaaaaatta ttcctcacagcagagtagctaaacctcatcaaagggaaaagattgaca aagaaatagagcttcacagaattcttcatcataagcatgtagtgcagt tttaccactacttcgaggacaaagaaaacatttacattctcttggaat actgcagtagaaggtcaatggctcatattttgaaagcaagaaaggtgt tgacagagccagaagttcgatactacctcaggcagattgtgtctggac tgaaataccttcatgaacaagaaatcttgcacagagatctcaaactag ggaacttttttattaatgaagccatggaactaaaagttggggacttcg gtctggcagccaggctagaacccttggaacacagaaggagaacgatat gtggtaccccaaattatctctctcctgaagtcctcaacaaacaaggac atggctgtgaatcagacatttgggccctgggctgtgtaatgtatacaa tgttactagggaggcccccatttgaaactacaaatctcaaagaaactt ataggtgcataagggaagcaaggtatacaatgccgtcctcattgctgg ctcctgccaagcacttaattgctagtatgttgtccaaaaacccagagg atcgtcccagtttggatgacatcattcgacatgacttttttttgcagg gcttcactccggacagactgtcttctagctgttgtcatacagttccag atttccacttatcaagcccagctaagaatttctttaagaaagcagctg ctgctctttttggtggcaaaaaagacaaagcaagatatattgacacac ataatagagtgtctaaagaagatgaagacatctacaagataggcatga tttgaaaaagacttcaataactcagcaacccagcaaacacaggacaga tgaggagctccagccacctaccaccacagttgccaggtctggaacacc cgcagtagaaaacaagcagcagattggggatgctatteggatgatagt cagagggactatggcagctgtagcagcagcagtgaatgccttgaagac agtaccatgggaagtgttgcagacacagtggcaagggttcttcgggga tgtctggaaaacatgccggaagctgattgcattcccaaagagcagctg agcacatcatttcagtgggtcaccaaatgggttgattactctaacaaa tatggctttgggtaccagctctcagaccacaccgtcggtgtccttttc aacaatggtgctcacatgagcctcatccagacaaaaaaacagttcact attacgcagagcttggccaatgctcagttttcccagcaacagatgctc ctgagcaatttattagtcaagtgacggtgctgaaatacttttctcatt acatggaggagaacctcatggatggtggagatctgcctagtgttactg atattcgaagaccteggctctacctccttcagtggctaaaatctgata aggccctaatgatgctctttaatgatggcacctttcaggtgaatttct accatgatcatacaaaaatcatcatctgtagccaaaatgaagaatacc ttctcacctacatcaatgaggataggatatctacaactttcaggctga caactctgctgatgtctggctgttcatcagaattaaaaaatcgaatgg aatatgccctgaacatgctcttacaaagatgtaactga Gene symbol: SRPK1 Entrez Gene ID: 6732 dna/ac: BC038292.1 sequence: (SEQ ID NO: 35) atggagcggaaagtgcttgcgctccaggcccgaaagaaaaggaccaag gccaagaaggacaaagcccaaaggaaatctgaaactcagcaccgaggc tctgctccccactctgagagtgatctaccagagcaggaagaggagatt ctgggatctgatgatgatgagcaagaagatcctaatgattattgtaaa ggaggttatcatcttgtgaaaattggagatctattcaatgggagatac catgtgatccgaaagttaggctggggacacttttcaacagtatggtta tcatgggatattcaggggaagaaatttgtggcaatgaaagtagttaaa agtgctgaacattacactgaaacagcactagatgaaatccggttgctg aagtcagttcgcaattcagaccctaatgatccaaatagagaaatggtt gttcaactactagatgactttaaaatatcaggagttaatggaacacat atctgcatggtatttgaagttttggggcatcatctgctcaagtggatc atcaaatccaattatcaggggcttccactgccttgtgtcaaaaaaatt attcagcaagtgttacagggtatgattatttacataccaagtgccgta tcacccacactgacattaaaccagagaacatcttattgtcagtgaatg agcagtacattcggaggctggctgcagaagcaacagaatggcagcgat ctggagctcctccgccttccggatctgcagtcagtactgctccccagc ctaaaccagctgacaaaatgtcaaagaataagaagaagaaattgaaga agaagcagaagcgccaggcagaattactagagaagcgaatgcaggaaa ttgaggaaatggagaaagagtcgggccctgggcaaaaaagaccaaaca agcaagaagaatcagagagtectgttgaaagaccatgaaagagaaccc acctaataaaatgacccaagaaaaacttgaagagtcaagtaccattgg ccaggatcaaacgcttatggaacgtgatacagagggtggtgcagcaga aattaattgcaatggagtgattgaagtcattaattatactcagaacag taataatgaaacattgagacataaagaggatctacataatgctaatga ctgtgatgtccaaaatttgaatcaggaatctagtttcctaagctccca aaatggagacagcagcacatctcaagaaacagactcttgtacacctat aacatctgaggtgtcagacaccatggtgtgccagtcttcctcaactgt aggtcagtcattcagtgaacaacacattagccaacttcaagaaagcat tcgggcagagataccctgtgaagatgaacaagagcaagaacataacgg accactggacaacaaaggaaaatccacggctggaaattttcttgttaa tccccttgagccaaaaaatgcagaaaagctcaaggtgaagattgctga ccttggaaatgcttgttgggtgcacaaacatttcactgaagatattca aacaaggcaatatcgttccttggaagttctaatcggatctggctataa tacccctgctgacatttggagcacggcatgcatggcctttgaactggc cacaggtgactatttgtttgaacctcattcaggggaagagtacactcg agatgaagatcacattgcattgatcatagaacttctggggaaggtgcc tcgcaagctcattgtggcaggaaaatattccaaggaatttttcaccaa aaaaggtgacctgaaacatatcacgaagctgaaaccttggggcctttt tgaggttctagtggagaagtatgagtggtcgcaggaagaggcagctgg cttcacagatttettactgcccatgttggagctgatccctgagaagag agccactgccgccgagtgtctccggcacccttggcttaactcctaa Gene symbol: SSA2 Entrez Gene ID: 6738 dna/ac: BC036658.2 sequence: (SEQ ID NO: 36) atggaggaatctgtaaaccaaatgcagccactgaatgagaagcagata gccaattctcaggatggatatgtatggcaagtcactgacatgaatcga ctacaccggttatatgatcggttctgaaggtgggacttattatatcaa agaacagaagttgggccttgaaaatgctgaagctttaattagattgat tgaagatggcagaggatgtgaagtgatacaagaaataaagtcatttag tcaagaaggcagaaccacaaagcaagagcctatgctctttgcacttgc catttgttcccagtgctccgacataagcacaaaacaagcagcatttaa agctgtttctgaagtttgtcgcattcctacccatctctttacttttat ccagtttaagaaagatctgaaggaaagcatgaaatgtggcatgtgggg tcgtgccctccggaaggctatagcggactggtacaatgagaaaggtgg catggcccttgctctggcagttacaaaatataaacagagaaatggctg gtctcacaaagatctattaagattgtcacatcttaaaccttccagtga aggacttgcaattgtgaccaaatatattacaaagggctggaaagaagt tcatgaattgtataaagaaaaagcactctctgtggagactgaaaaatt attaaagtatctggaggctgtagagaaagtgaagcgcacaagagatga gctagaagtcattcatctaatagaagaacatagattagttagagaaca tcttttaacaaatcacttaaagtctaaagaggtatggaaggctttgtt acaagaaatgccgcttactgcattactaaggaatctaggaaagatgac tgctaattcagtacttgaaccaggaaattcagaagtatctttagtatg tgaaaaactgtgtaatgaaaaactattaaaaaaggctcgtatacatcc atttcatattttgatcgcattagaaacttacaagacaggtcatggtct cagagggaaactgaagtggcgccctgatgaagaaattttgaaagcatt ggatgctgctttttataaaacatttaagacagttgaaccaactggaaa acgtttcttactagctgttgatgtcagtgcttctatgaaccaaagagt tttgggtagtatactcaacgctagtacagttgctgcagcaatgtgcat ggttgtcacacgaacagaaaaagattatatgtagttgattttccgatg aaatggtaccatgtccagtgactacagatatgaccttacaacaggttt taatggctatgagtcagatcccagcaggtggaactgattgctctcttc caatgatctgggctcagaagacaaacacacctgctgatgtatcattgt attcactgataatgagacattgctggaggtgtccatcctgctattgct ctgagggagtatcgaaagaaaatggatattccagctaaattgattgtt tgtggaatgacatcaaatggtttcaccattgcagacccagatgataga ggcatgttggatatgtgcggattgatactggagctctggatgtaattc gaaatttcacattagatatgatttaa Gene symbol: SSNA1 Entrez Gene ID: 8636 dna/ac: BC000864.1 sequence: (SEQ ID NO: 37) atgacccagcagggcgcggcgctgcagaactacaacaacgagctggtc aagtgcatagaggagctgtgccagaagcgggaggagctgtgccggcag atccaggaggaggaggacgagaagcagcggctgcagaatgaggtgagg cagctgacagagaagctggcccgcgtcaacgagaacctggcacgcaag attgcctctcgcaacgagttcgaccggaccatcgcggagacggaggcc gcctacctcaagatcctggagagctcccagactttgctcagcgttctc aagagggaagctgggaacctgaccaaggctacagccccagaccagaaa agtagcggcggcagggacagctga Gene symbol: STAU1 Entrez Gene ID: 6780 dna/ac: BC050432.1 sequence: (SEQ ID NO: 38) atgtctcaagttcaagtgcaagttcagaacccatctgctgctctctca gggagccaaatactgaacaagaaccagtctcttctctcacagcctttg atgagtattccttctactactagctctctgccctctgaaaatgcaggt agacccattcaaaactctgattaccctctgcatctattacatccacca gtgcagctgcagaaagcataacccctactgtagaactaaatgcactgt gcatgaaacttggaaaaaaaccaatgtataagcctgttgacccttact ctcggatgcagtccacctataactacaacatgagaggaggtgcttatc ccccgaggtacttttacccatttccagttccacctttactttatcaag tggaactttctgtgggaggacagcaatttaatggcaaaggaaagacaa gacaggctgcgaaacacgatgctgctgccaaagcgttgaggatcctgc agaatgagcccctgccagagaggctggaggtgaatggaagagaatccg aagaagaaaatctcaataaatctgaaataagtcaagtgfttgagattg cacttaaacggaacttgcctgtgaatttcgaggtggcccgggagagtg gcccaccccacatgaagaactttgtgaccaaggtttcggttggggagt ttgtgggggaaggtgaagggaaaagcaagaagatttcaaagaaaaatg ccgccatagctgttcttgaggagctgaagaagttaccgcccctgcctg cagttgaacgagtaaagcctagaatcaaaaagaaaacaaaacccatag tcaagccacagacaagcccagaatatggccaggggatcaatccgatta gccgactggcccagatccagcaggcaaaaaaggagaaggagccagagt acacgctcctcacagagcgaggcctcccgcgccgcagggagtttgtga tgcaggtgaaggttggaaaccacactgcagaaggaacgggcaccaaca agaaggtggccaagcgcaatgcagccgagaacatgctggagatccttg gtttcaaagtcccgcaggcgcagcccaccaaacccgcactcaagtcag aggagaagacacccataaagaaaccaggggatggaagaaaagtaacct tttttgaacctggctctggggatgaaaatgggactagtaataaagagg atgagttcaggatgccttatctaagtcatcagcagctgcctgctggaa ttcttcccatggtgcccgaggtcgcccaggctgtaggagttagtcaag gacatcacaccaaagattttaccagggcagctccgaatcctgccaagg ccacggtaactgccatgatagcccgagagttgttgtatgggggcacct cgcccacagccgagaccattttaaagaataacatctcttcaggccacg taccccatggacctctcacgagaccctctgagcaactggactatcttt ccagagtccagggattccaggttgaatacaaagacttccccaaaaaca acaagaacgaatttgtatctcttatcaattgctcctctcagccacctc tgatcagccatggtatcggcaaggatgtggagtcctgccatgatatgg ctgcgctgaacatcttaaagttgctgtctgagttggaccaacaaagta cagagatgccaagaacaggaaacggaccaatgtctgtgtgtgggaggt gctga Gene symbol: STK11 Entrez Gene ID: 6794 dna/ac: BC007981.2 sequence: (SEQ ID NO: 39) atggaggtggtggacccgcagcagctgggcatgttcacggagggcgag ctgatgtcggtgggtatggacacgttcatccaccgcatcgactccacc gaggtcatctaccagccgcgccgcaagcgggccaagctcatcggcaag tacctgatgggggacctgctgggggaaggctcttacggcaaggtgaag gaggtgctggactcggagacgctgtgcaggagggccgtcaagatcctc aagaagaagaagttgcgaaggatccccaacggggaggccaacgtgaag aaggaaattcaactactgaggaggttacggcacaaaaatgtcatccag ctggtggatgtgttatacaacgaagagaagcagaaaatgtatatggtg atggagtactgcgtgtgtggcatgcaggaaatgctggacagcgtgccg gagaagcgtttcccagtgtgccaggcccacgggtacttctgtcagctg attgacggcctggagtacctgcatagccagggcattgtgcacaaggac atcaagccggggaacctgctgctcaccaccggtggcaccctcaaaatc tccgacctgggcgtggccgaggcactgcacccgttcgcggcggacgac acctgccggaccagccagggctccccggctttccagccgcccgagatt gccaacggcctggacaccttctccggcttcaaggtggacatctggtcg gctggggtcaccctctacaacatcaccacgggtctgtaccccttcgaa ggggacaacatctacaagttgtttgagaacatcgggaaggggagctac gccatcccgggcgactgtggccccccgctctctgacctgctgaaaggg atgcttgagtacgaaccggccaagaggttctccatccggcagatccgg cagcacagctggttccggaagaaacatcctccggctgaagcaccagtg cccatcccaccgagcccagacaccaaggaccggtggcgcagcatgact gtggtgccgtacttggaggacctgcacggcgcggacgaggacgaggac ctcttcgacatcgaggatgacatcatctacactcaggacttcacggtg cccggacaggtcccagaagaggaggccagtcacaatggacagcgccgg ggcctccccaaggccgtgtgtatgaacggcacagaggcggcgcagctg agcaccaaatccagggcggagggccgggcccccaaccctgcccgcaag gcctgctccgccagcagcaagatccgccggctgtcggcctgcaagcag cagtga Gene symbol: TOM1 Entrez Gene ID: 10043 dna/ac: BC046151.1 sequence: (SEQ ID NO: 40) atggactttctcctggggaacccgttcagctctccagtgggacagcgc atcgagaaagccacagatggctccctgcagagcgaggactgggccctc aacatggagatctgcgacatcatcaacgagacggaggaaggtcccaaa gatgccctccgagcagtaaagaagagaatcgtggggaataagaacttc cacgaggtgatgctggctctcacagtcttagaaacctgtgtcaagaac tgcgggcaccgcnccacgtgctggtggccagccaggacncgtggagag tgtgctggtgaggaccatcctgcccaagaacaacccacccaccatcgt gcatgacaaagtgctcaacctcatccagtcctgggctgacgcgttccg cagctcgcccgatctgacaggtgtggtcaccatctatgaggacctgcg gaggaaaggcctggagttccccatgactgacctggacatgctgtcacc catccacacaccccagaggaccgtgttcaactcagagacacaatcagg acaggattctgtgggcactgactccagccagcaagaggactctggcca gcatgctgcccctctgcccgccccgcccatactctccggtgacacgcc catagcaccaaccccggaacagattgggaagctgcgcagtgagctgga gatggtgagtgggaacgtgagggtgatgtcggagatgctgacggagct ggtgcccacccaggccgagcccgcagacctggagctgctgcaggagct caaccgcacgtgccgagccatgcagcagcgggtcctggagctcatccc tcagatcgccaatgagcagctgacagaggagctgctcatcgtcaatga caatctcaacaatgtgttcctgcgccatgaacggtttgaacggttccg aacaggccagaccaccaaggccccaagtgaggccgagccggcagctga cctgatcgacatgggccctgacccagcagccaccggcaacctctcatc ccagctggcaggaatgaacctgggctccagcagtgtgagagctggcct gcagtctctggaggcctctggtcgactggaagatgagtttgacatgtt tgcgctgacacggggcagctcactggctgaccaacggaaagaggtaaa atacgaagccccccaagcaacagacggcctggctggagccctggacgc ccggcagcagagcactggcgcgatcccagtcacccaggcctgcctcat ggaggacatcgagcagtggctgtccactgacgtgggtaatgatgcgga agagcctaagggggtcaccagcgaaggtaaatttgacaaattcctgga agaacgggccaaagccgcggaccgattgcccaacctctccagcccctc agctgaggggcccccgggtcccccatctggcccagcgccccggaagaa gacccaggagaaagatgatgacatgctgtttgccttatga Gene symbol: TXNL2 Entrez Gene ID: 10539 dna/ac: BC005289 sequence: (SEQ ID NO: 41) atggcggcgggggcggctgaggcagctgtagcggccgtggaggaggtc ggctcagccgggcagtttgaggagctgctgcgcctcaaagccaagtcc ctccttgtggtccatttctgggcaccatgggctccacagtgtgcacag atgaacgaagttatggcagagttagctaaagaactccctcaagtttca tttgtgaagttggaagctgaaggtgttcctgaagtatctgaaaaatat gaaattagctctgttcccacttttctgtttttcaagaattctcagaaa atcgaccgattagatggtgcacatgccccagagttgaccaaaaaagtt cagcgacatgcatctagtggctccttcctacccagcgctaatgaacat cttaaagaagatctcaaccttcgcttgaagaaattgactcatgctgcc ccctgcatgctgtttatgaaaggaactcctcaagaaccacgctgtggt ttcagcaagcagatggtggaaattcttcacaaacataatattcagttt agcagttttgatatcttctcagatgaagaggttcgacagggactcaaa gcctattccagttggcctacctatcctcagctctatgtttctggagag ctcataggaggacttgatataattaaggagctagaagcatctgaagaa ctagatacaatttgtcccaaagctcccaaattagaggaaaggctcaaa gtgctgacaaataaagcttctgtgatgctctttatgaaaggaaacaaa caggaagcaaaatgtggattcagcaaacaaattctggaaatactaaat agtactggtgttgaatatgaaacattcgatatattggaggatgaagaa gttcggcaaggattaaaagcttactcaaattggccaacataccctcag ctgtatgtgaaaggggagctggtgggaggattggatattgtgaaggaa ctgaaagaaaatggtgaattgctgcctatactgagaggagaaaattaa Gene symbol: TXNRD1 Entrez Gene ID: 7296 dna/ac: BC018122.1 sequence: (SEQ ID NO: 42) atgaacggccctgaagatcttcccaagtcctatgactatgaccttatc atcattggaggtggctcaggaggtctggcagctgctaaggaggcagcc caatatggcaagaaggtgatggtcctggactttgtcactcccacccct cttggaactagatggggtcttggaggaacatgtgtgaatgtgggttgc atacctaaaaaactgatgcatcaagcagctttgttaggacaagccctg caagactctcgaaattatggatggaaagtcgaggagacagttaagcat gattgggacagaatgatagaagctgtacagaatcacattggctctttg aattggggctaccgagtagctctgcgggagaaaaaagtcgtctatgag aatgcttatgggcaatttattggtcctcacaggattaaggcaacaaat aataaaggcaaagaaaaaatttattcagcagagagatttctcattgcc actggtgaaagaccacgttacttgggcatccctggtgacaaagaatac tgcatcagcagtgatgatcttttctccttgccttactgcccgggtaag accctggttgttggagcatcctatgtcgctttggagtgcgctggattt cttgctggtattggtttagacgtcactgttatggttaggtccattctt cttagaggatttgaccaggacatggccaacaaaattggtgaacacatg gaagaacatggcatcaagtttataagacagttcgtaccaattaaagtt gaacaaattgaagcagggacaccaggccgactcagagtagtagctcag tccaccaatagtgaggaaatcattgaaggagaatataatacggtgatg ctggcaataggaagagatgcttgcacaagaaaaattggcttagaaacc gtaggggtgaagataaatgaaaagactggaaaaatacctgtcacagat gaagaacagaccaatgtgccttacatctatgccattggcgatatattg gaggataaggtggagctcaccccagttgcaatccaggcaggaagattg ctggctcagaggctctatgcaggttccactgtcaagtgtgactatgaa aatgttccaaccactgtatttactcctttggaatatggtgcttgtggc ctttctgaggagaaagctgtggagaagtttggggaagaaaatattgag gtttaccatagttacttttggccattggaatggacgattccgtcaaga gataacaacaaatgttatgcaaaaataatctgtaatactaaagacaat gaacgtgttgtgggctttcacgtactgggtccaaatgctggagaagtt acacaaggctttgcagctgcgctcaaatgtggactgaccaaaaagcag ctggacagcacaattggaatccaccctgtctgtgcagaggtattcaca acattgtctgtgaccaagcgctctggggcaagcatcctccaggctggc tgctgaggttaa Gene symbol: VCL Entrez Gene ID: 7414 dna/ac: BC039174.1 sequence: (SEQ ID NO: 43) atgccagtgtttcatacgcgcacgatcgagagcatcctggagccggtg gcacagcagatctcccacctggtgataatgcacgaggagggcgaggtg gacggcaaagccattcctgacctcaccgcgcccgtggccgccgtgcag gcggccgtcagcaacctcgtccgggttggaaaagagactgttcaaacc actgaggatcagattttgaagagagatatgccaccagcatttattaag gttgagaatgcttgcaccaagcttgtccaggcagctcagatgcttcag tcagacccttactcagtgcctgctcgagattatctaattgatgggtca aggggcatcctctctggaacatcagacctgctccttaccttcgatgag gctgaggtccgtaaaattattagagtttgcaaaggaattttggaatat cttacagtggcagaggtggtggagactatggaagatttggtcacttac acaaagaatcttgggccaggaatgactaagatggccaagatgattgac gagagacagcaggagctcactcaccaggagcaccgagtgatgttggtg aactcgatgaacaccgtgaaagagttgctgccagttctcatttcagct atgaagatttttgtaacaactaaaaactcaaaaaaccaaggcatagag gaagctttaaaaaatcgcaattttactctagaaaaaatgagtgctgaa attaatgagataattcgtgtgttacaactcacctctgggatgaagatg cctgggccagcaaggacactgaagccatgaagagagcattggcctcca tagactccaaactgaaccaggccaaaggttggctccgtgaccctagtg cctccccaggggatgctggtgagcaggccatcagacagatcttagatg aagctggaaaagttggtgaactctgtgcaggcaaagaacgcagggaga ttctgggaacttgcaaaatgctagggcagatgactgatcaagtggctg acctccgtgccagaggacaaggatcctcaccggtggccatgcagaaag ctcagcaggtatctcagggtctggatgtgctcacagcaaaagtggaaa atgcagctcgcaagctggaagccatgaccaactcaaagcagagcattg caaagaagatcgatgctgctcagaactggcttgcagatccaaatggtg gaccggaaggagaagagcagattcgaggtgctttggctgaagctcgga aaatagcagaattatgtgatgatcctaaagaaagagatgacattctac gttcccttggggaaatatctgctctgacttctaaattagcagatctac gaagacaggggaaaggagattctccagaggctcgagccttggccaaac aggtggccacggccctgcagaacctgcagaccaaaaccaaccgggctg tggccaacagcagaccggccaaagcagctgtacaccttgagggcaaga ttgagcaagcacagcggtggattgataatcccacagtggatgaccgtg gagtcggtcaggctgccatccgggggcttgtggccgaagggcatcgtc tggctaatgttatgatggggccttatcggcaagatcttctcgccaagt gtgaccgagtggaccagctgacagcccagctggctgacctggctgcca gaggggaaggggagagtcctcaggcacgagcacttgcatctcagctcc aagactccttaaaggatctaaaagctcggatgcaggaggccatgactc aggaagtgtcagatgttttcagcgataccacaactcccatcaagctgt tggcagtggcagccacggcgcctcctgatgcgcctaacagggaagagg tatttgatgagagggcagctaactttgaaaaccattcaggaaagcttg gtgctacggccgagaaggcggctgcggttggtactgctaataaatcaa cagtggaaggcattcaggcctcagtgaagacggcccgagaactcacac cccaggtggtctcggctgctcgtatcttacttaggaaccctggaaatc aagctgcttatgaacattttgagaccatgaagaaccagtggatcgata atgttgaaaaaatgacagggctggtggacgaagccattgataccaaat ctctgttggatgcttcagaagaagcaattaaaaaagacctggacaagt gcaagglagctatggccaacattcagcctcagatgctggttgctgggg caaccagtattgctcgtcgggccaaccggatcctgctggtggctaaga gggaggtggagaattccgaggatcccaagttccgtgaggctgtgaaag ctgcctctgatgaattgagcaaaaccatctccccgatggtgatggatg caaaagctgtggctggaaacatttccgaccctggactgcaaaagagct tcctggactcaggatatcggatcctgggagctgtggccaaggtcagag aagccttccaacctcaggagcctgacttcccgccgcctccaccagacc ttgaacaactccgactaacagatgagcttgctcctcccaaaccacctc tgcctgaaggtgaggtccctccacctaggcctccaccaccagaggaaa aggatgaagagttccctgagcagaaggccggggaggtgattaaccagc caatgatgatggctgccagacagctccatgatgaagctcgcaaatggt ccagcaagggcaatgacatcattgcagcagccaagcgcatggctctgc tgatggctgagatgtctcggctggtaagagggggcagtggtaccaagc gggcactcattcagtgtgccaaggacatcgccaaggcctcagatgagg tgactcggttggccaaggaggttgccaagcagtgcacagataaacgga ttagaaccaacctcttacaggtatgtgagcgaatcccaaccataagca ccagctcaaaatcctgtccacagtgaaggccaccatgctgggccggac caacatcagtgatgaggagtctgagcaggccacagagatgctggttca caatgcccagaacctcatgcagtctgtgaaggagactgtgcgggaagc tgaagctgcttcaatcaaaattcgaacagatgctggatttacactgcg ctgggttagaaagactccctggtaccagtag Gene symbol: ZMAT2 Entrez Gene ID: 153527 dna/ac: BC056668.1 sequence: (SEQ ID NO: 44) atggcgtcgggcagcgggacaaaaaacttggactttcgccgaaagtgg gacaaagatgaatatgagaaactcgccgagaagaggctcacggaagag agagaaaagaaagatggaaaaccagtgcagcctgtcaagcgagagctt ttacggcatagggactacaaggtggacgggaatccaagcttgggaaga caattgtcattaccaagacaacccctcaatctgagatgggaggatatt actgcaatgtctgtgactgtgtggtgaaggactccatcaactttctgg atcacattaatggaaagaaacatcagagaaacctgggcatgtctatgc gtgtggaacgttccaccctggatcaggtgaagaaacgttttgaggtca acaagaagaagatggaagagaagcagaaggattatgattttgaggaaa ggatgaaggagctcagagaagaggaggaaaaggccaaagcgtacaaga aagagaaacagaaggagaagaaaaggagggctgaggaggacttgacat ttgaggaggacgatgagatggcagctgtgatgggcttctctggctttg gttccaccaagaagagttactga Gene symbol: ZNF38 Entrez Gene ID: 7589 dna/ac: BC047309.1 sequence: (SEQ ID NO: 45) atgatgaccaaggtactaggcatggccccagttctgggccctaggcct ccacaggagcaggtggggcctctgatggtaaaagtcgaggagaaagaa gagaaaggcaagtaccttcctagcctggagatgttccgccagcgcttc aggcagtttgggtaccatgatacccctggaccccgagaggccctgagc caactccgggtgctctgctgtgagtggctgaggcccgagatccacacc aaggagcagatcctggagctactggtgctggagcagttcctgaccatc ctgccccaggagaccaggcctgggtgcaggagcattgcccggagagcg ctgaagaggctgtcactctcctcgaagatctggagcgggaactggatg agccaggacaccaggtctcaactcctccaaacgaacagaaaccggtgt gggagaagatatcctcctcaggaactgcaaaggaatccccgagcagca tgcagccacagcccttggagaccagtcacaaatacgagtcttgggggc ccctgtacatccaagagtctggtgaggagcaggagttcgctcaagatc caagaaaggtccgagattgcagattgagtacccagcacgaggaatcag cagatgagcagaaaggactgaagcagaggggctcaaaggggatataat ttctgtgattatcgccaataaacctgaggccagcttagagaggcagtg cgtaaaccttgaaatgaaaaaggaacaaaaccccctatcaagaggcag gctccaagaaaggtagagaatcagttcctactaaacctaccccaggag agagacgttatatatgtgctgaatgtggcaaagcctttagtaatagct caaatctcaccaaacacaggagaacacacactggggagaaaccttacg tgtgcaccaagtgtgggaaagctttcagccacagctcaaacctcacca ccactacagaacacacttggtggaccggccetatgactgtaagtgtgg aaaagcttttgggcagagctcagaccttcttaaacatcagagaatgca cacagaagaggcgccatatcagtgcaaagattgtggcaaggctttcag cgggaaaggcagcctcattcgtcactatcggatccacactggggagaa gccttatcagtgtaacgaatgtgggaagagcttcagtcagcatgcggg cctcagacccaccagagactccacaccggagagaagccatataagtgt aaggagtgtgggaaagccttcaaccacagctccaacttcaataaacac cacagaatccacaccggggaaaagccctactggtgtcatcactgtgga aagaccttctgtagcaagtccaatctttccaaacatcagcgagtccac actggagagggagaagcaccgtaa 

1. A method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises: (a) ZMAT2 and the other 31 auto-antigens listed in Table 2, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced.
 2. A method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises: (a) the auto-antigens listed in Table 1, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced.
 3. A method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises: (a) the auto-antigens listed in Table
 4. (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced.
 4. A method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises: (a) the auto-antigens listed in Table 3, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 10, 15, 20 or 25 or more auto-antigens are deleted or replaced.
 5. A method of diagnosing a patient as having or being predisposed to developing lupus, comprising detecting in a sample taken from said patient one or more auto-antibodies that bind an auto-antigen selected from the group consisting of the auto-antigens listed in Table 3, wherein the presence of said one or more auto-antibodies, or an increase in the concentration of said one or more auto-antibodies, indicates that the patient suffers from or is predisposed to develop lupus.
 6. A method useful in diagnosing a patient as having or being predisposed to developing lupus, comprising testing a sample from the patient against a panel of auto-antigens to detect the presence, absence or a modulated level of auto-antibodies in the sample, wherein the panel of auto-antigens comprises at least 10 auto-antigens selected from Table
 3. 7. A method according to claim 6, wherein the at least 10 auto-antigens are selected from Table
 1. 8. A method according to claim 6, wherein the at least 10 auto-antigens are selected from Table
 2. 9. A method according to claim 6, wherein the panel comprises at least 20 auto-antigens and/or wherein the panel comprises ZMAT2.
 10. A method according to claim 3, wherein the panel comprises one or more additional auto-antigens selected from the group consisting of the auto-antigens of Table
 5. 11. The method of claim 1 wherein detecting the presence or an increased level of at least two auto-antibodies provides information useful for diagnosis.
 12. The method of claim 11, wherein detecting the presence of an increased level of at least three, preferably at least four and more preferably at least five auto-antibodies provides information useful for diagnosis.
 13. The method of claim 1, wherein the sample is a body fluid.
 14. The method of claim 13, wherein the body fluid is selected from blood, serum, plasma, saliva, lymphatic fluid, wound secretion, urine, faeces, mucus or cerebrospinal fluid (CSF).
 15. (canceled)
 16. A method of monitoring the progression or regression of lupus, or monitoring the progression to a flare of the disease or transition from a flare into remission, comprising: (a) performing the method of claim 1, (b) repeating step (a) using a second sample obtained from the patient at a time later than the first sample; (c) comparing the auto-antibodies detected in said first and second samples; wherein a change in the level or presence or absence of the auto-antibodies detected between the first and second samples indicates the progression or regression of lupus, or the progression to a flare, or transition from a flare into remission.
 17. A method of monitoring the efficacy in a patient of a therapeutic agent to lupus, comprising: (a) performing the method of claim 1, (b) administering the therapeutic agent to the patient; (c) performing the method of claim 1 using a second sample from the patient taken after the administration of the therapeutic agent; (d) comparing the auto-antibodies detected in said first and second samples; wherein a change in the level or presence or absence of the auto-antibodies detected in the first and second sample is indicative of the efficacy of the therapeutic agent to lupus.
 18. A method of diagnosing a patient as having or being predisposed to developing lupus, comprising detecting in a sample taken from said patient one or more auto-antibodies that bind an auto-antigen selected from: a) the group consisting of ZMAT2 (Zinc finger, matrin type 2); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Romeo box B6, transcript variant 2); KRT8 (Keratin 8); LIN28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class H)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single, stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); and ZNF38 (Zinc finger protein 38), or b) the group consisting of ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); KRT8 (Keratin 8); LIN28 Lin-8 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1); STAU1 (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); and ZNF38 (Zinc finger protein 38), wherein the presence of said one or more auto-antibodies, or an increase in the concentration of said one or more auto-antibodies, indicates that the patient suffers from or is predisposed to develop lupus.
 19. The method of claim 18, wherein the presence of auto-antibodies to at least two of the auto-antigens are detected.
 20. A method of diagnosing a patient as having or being predisposed to developing lupus, comprising detecting in a sample from said patient auto-antibodies to a panel comprising 5 or more auto-antigens selected from: a) the group consisting of ZMAT2 (Zinc finger, matrin type 2); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); LIN28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron
 1. telomeric); SRPK1 (SFRS protein kinase 1): SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase
 1. transcript variant 5); and ZNF38 (Zinc finger protein 38); or b) the group consisting of ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); KRT8 (Keratin 8); LIN28 Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1); STAU1(Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); ZNF38 (Zinc finger protein 38). wherein the presence or separately a significant increase in the concentration of said auto-antibodies in the sample to substantially all members of the panel indicates that the patient suffers from or is predisposed to develop lupus.
 21. The method of claim 20, wherein the panel comprises 10 or more of said auto-antigens. 22-39. (canceled)
 40. The method of claim 1, wherein the sample is a body fluid.
 41. The method of claim 40, wherein the body fluid is selected from blood, serum, plasma, saliva, lymphatic fluid, wound secretion, urine, faeces, mucus or cerebrospinal fluid (CSF).
 42. The method of claim 41, wherein the sample is serum or plasma.
 43. A diagnostic kit for use in diagnosing a patient as having or being predisposed to developing lupus, said kit comprising one or more auto-antigens selected from: (a) the group consisting of ZMAT2 (Zinc finger, matrin type 2); BPY2IP1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); LIN28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein
 12. transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); ZNF38 (Zinc finger protein 38); and (b) the group consisting of the auto-antigens of ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12; 16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); KRT8 (Keratin 8); LIN28 Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1); STAU1 (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); ZNF38 (Zinc finger protein 38), or (c) the group consisting of the auto-antigens of Table 3; which one or more auto-antigens are immobilised on a substrate.
 44. A diagnostic kit for use in diagnosing a patient as having or being predisposed to developing lupus, said kit comprising a panel comprising 5 or more auto-antigens selected from: (a) the group consisting of ZMAT2 (Zinc finger, matrin type 2); BPY21P1 (MAP1S protein); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); E1B-AP5 (E1B-55 kDa-associated protein 5); FUS (Fusion (involved in t(12;16) in malignant liposarcoma)); HAGH (Hydroxyacylglutathione hydrolase); HMG20B (High-mobility group 20B); HOXB6 (Homeo box B6, transcript variant 2); KRT8 (Keratin 8); LIN28 (Lin-28 homolog (C. elegans)); NDUFV3 (NADH dehydrogenase (ubiquinone) flavoprotein 3, 10 kDa); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); RAB11FIP3 (RAB11 family interacting protein 3 (class II)); RAN (RAN, member RAS oncogene family); RARA (Retinoic acid receptor alpha); RBMS1 (RNA binding motif, single stranded interacting protein 1, transcript variant); RDBP (RD RNA binding protein); RNF12 (Ring finger protein 12, transcript variant 1); RUFY1 (RUN and FYVE domain containing 1); SMN1 (Survival of motor neuron 1, telomeric); SRPK1 (SFRS protein kinase 1); SSNA1 (Sjogren's syndrome nuclear autoantigen 1); STAU (Staufen, RNA binding protein (Drosophila), transcript variant T3); STK11 (Serine/threonine kinase 11 (Peutz-Jeghers syndrome)); TOM1 (Target of myb1 (chicken)); TXNL2 (Thioredoxin-like, clone MGC:12349); TXNRD1 (Thioredoxin reductase 1, transcript variant 5); and ZNF38 (Zinc finger protein 38); (b) the group consisting of the auto-antigens ZMAT2 (Zinc finger, matrin type 2); ASPSCR1 (alveolar soft part sarcoma chromosome region, candidate 1); CEBPG (CCAAT/enhancer binding protein (C/EBP), gamma); DDX55 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 55); DOM3Z (dom-3 homolog Z (C. elegans)); E1B-AP5 (E1B-55 kDa-associated protein 5) FUS (Fusion (involved in t(12; 16) in malignant liposarcoma)); HMG20B (High-mobility group 20B); KRT8 (Keratin 8); LIN28 Lin-28 homolog (C. elegans)); LNX (ligand of numb-protein X); MAP1S (MAP1S protein); PABPC1 (Poly(A) binding protein, cytoplasmic 1); PHLDA1 (Pleckstrin homology-like domain, family A, member 1); PIAS2 (Msx-interacting-zinc finger, transcript variant alpha); PRKCBP1 (protein kinase C binding protein 1); PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator); RARA (Retinoic acid receptor, alpha); RDBP (RD RNA binding protein); RPL30 (ribosomal protein L30); RPL31 (ribosomal protein L31); SNK (serum-inducible kinase); SRPK1 (SFRS protein kinase 1); STAU1 (Staufen, RNA binding protein (Drosophila), transcript variant T3); TXNL2 (Thioredoxin-like, clone MGC:12349); VCL (vinculin); and ZNF38 (Zinc finger protein 38); or (c) the group consisting of the auto-antigens of Table 3; wherein the auto-antigens are immobilised on a substrate. 45-59. (canceled)
 60. A panel of auto-antigens, wherein said panel comprises or consists of: (i) (a) the auto-antigens listed in Table 2, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 6, 7, 8, 9 or 10-ere or more auto-antigens are deleted or replaced; (ii) (a) the auto-antigens listed in Table 1, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced; (iii) (a) the auto-antigens listed in Table 4, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 6, 7, 8, 9 or 10 or more auto-antigens are deleted or replaced; or (iv) (a) the auto-antigens listed in Table 3, (b) the panel of (a) in which up to 5 auto-antigens are deleted or replaced, or (c) the panel of (a) in which up to 10, 15, 20 or 25 or more auto-antigens are deleted or replaced.
 61. A panel according to claim 60 (i)(b), claim 60 (i)(c), claim 60 (ii)(b), claim 60 (ii)(c), claim 60 (iii)(b), claim 60 (iii)(c), claim 60 (iv)(b) or claim 60 (iv)(c) or a method according to claim 60 which method specifies a panel, wherein the panel comprises ZMAT2.
 62. A panel according to claim 60 wherein the auto-antigens are variants.
 63. A method according to claim 1 wherein the absence of said auto-antibodies in the sample indicates that the patient does not have or is not predisposed to developing lupus thereby providing a negative diagnosis.
 64. A method according to claim 18 wherein the method further comprises detecting in a sample taken from said patient one or more auto-antibodies that bind an auto-antigen selected from the group consisting of: BANK1 (B-cell scaffold protein with ankyrin repeats 1); IFI16 (Interferon, gamma-inducible protein 16); PSME3 (Proteasome (prosome, macropain) activator subunit 3 (PA28 gamma; Ki), transc); RALBP1 (RalA-binding protein 1); and SSA2 (Sjogren syndrome antigen A2 (60 kDa, ribonucleoprotein autoantigen SS-A/Ro)). 